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Juvenile hormones (JHs) play a crucial role in regulating development and reproduction in insects. Most insects predominantly synthesize JH III, which typically involves esterification followed by epoxidation, lepidopteran insects use a pathway of epoxidation followed by esterification. Although hemipteran insects have JH III and JH skipped bisepoxide III (JH SB3), the synthesis pathway and key epoxidases remain unclear. This study was conducted on Aphis craccivora, and demonstrated that corpora allata, microsomes, Ac‐CYP15C1, and Ac‐JHAMT catalyze JH III production in vitro, establishing the pathway of epoxidation followed by esterification. These findings were further confirmed through RNA interference and molecular docking. The presence of JH III and JH SB3 in A. craccivora was identified, and their synthesis pathway was elucidated as follows: Ac‐CYP15C1 oxidizes farnesic acid to JH A, followed by methylation to JH III by Ac‐JHAMT, possibly providing an epoxidation site on the second carbon for JH SB3. This alteration may significantly contribute to the differentiation and functional diversification of JH types in insects. [ABSTRACT FROM AUTHOR]

Avian influenza (AI), caused by H9N2 subtype avian influenza virus (AIV), poses a serious threat to poultry farming and public health due to its transmissibility and pathogenicity. The PB2 protein is a major component of the viral RNA polymerase complex. It is of great importance to identify the antigenic determinants of the PB2 protein to explore the function of the PB2 protein. In this study, the PB2 sequence of H9N2 subtype AIV, from 1090 to 1689 bp, was cloned and expressed. The recombinant PB2 protein with cutting gel was used to immunize BALB/c mice. After cell fusion, the hybridoma cell lines secreting monoclonal antibodies (mAbs) targeting the PB2 protein were screened by indirect ELISA and western blotting, and the antigenic epitopes of mAbs were identified by constructing truncated overlapping fragments in the PB2 protein of H9N2 subtype AIV. The results showed that three hybridoma cell lines (4B7, 4D10, and 5H1) that stably secreted mAbs specific to the PB2 protein were screened; the heavy chain of 4B7 was IgG2α, those of 4D10 and 5H1 were IgG1, and all three mAbs had kappa light chain. Also, the minimum B-cell epitope recognized was 475LRGVRVSK482 and 528TITYSSPMMW537. Homology analysis showed that these two epitopes were conserved among the different subtypes of AIV strains and located on the surface of the PB2 protein. The above findings provide an experimental foundation for further investigation of the function of the PB2 protein and developing monoclonal antibody-based diagnostic kits. [ABSTRACT FROM AUTHOR]

Cysteine plays a pivotal role in the sulfur metabolism network of plants, intimately influencing the conversion rate of organic sulfur and the plant's capacity to withstand abiotic stresses. In tea plants, the serine acetyltransferase (SAT) genes emerge as a crucial regulator of cysteine metabolism, albeit with a notable lack of comprehensive research. Utilizing Hidden Markov Models, we identified seven CssSATs genes within the tea plant genome. The results of the bioinformatics analysis indicate that these genes exhibit an average molecular weight of 33.22 kD and cluster into three distinct groups. Regarding gene structure, CssSAT1 stands out with ten exons, significantly more than its family members. In the promoter regions, cis-acting elements associated with environmental responsiveness and hormone induction predominate, accounting for 34.4% and 53.1%, respectively. Transcriptome data revealed intricate expression dynamics of CssSATs under various stress conditions (e.g., PEG, NaCl, Cold, MeJA) and their tissue-specific expression patterns in tea plants. Notably, qRT-PCR analysis indicated that under salt stress, CssSAT1 and CssSAT3 expression levels markedly increased, whereas CssSAT2 displayed a downregulatory trend. Furthermore, we cloned CssSAT1-CssSAT3 genes and constructed corresponding prokaryotic expression vectors. The resultant recombinant proteins, upon induction, significantly enhanced the NaCl tolerance of Escherichia coli BL21, suggesting the potential application of CssSATs in bolstering plant stress resistance. These findings have enriched our comprehension of the multifaceted roles played by CssSATs genes in stress tolerance mechanisms, laying a theoretical groundwork for future scientific endeavors and research pursuits. [ABSTRACT FROM AUTHOR]

Harpins play a key role in inducing disease resistance in crops, and identifying their core functional regions and establishing a system for their efficient expression would be very valuable. In this study, large amounts of soluble fusion proteins of harpin HrpZ and its subpeptides were obtained via the optimized induction conditions (28 ℃C with 0.5 mmol.L-1 IPTG for 6 h) in Escherichia coli BL21 (DE3). Hypersensitive response (HR) assays demonstrated that the C-terminal 66 aa of HrpZ (HrpZ_C_2_2) elicited a strong HR in tobacco (Nicotiana benthamiana) and grape (Flame Seedless) leaves. Additionally, treatment with HrpZ, and particularly HrpZ_C_2_2, significantly reduced the disease incidence and severity index of field vine leaves and those inoculated with downy mildew. The determination of the physiological parameters indicated that HrpZ, and especially HrpZ_C_2_2, improved the photosynthesis- and chlorophyll fluorescence-related parameters, enhanced the activity of defense-related enzymes, including SOD, POD, CAT and PAL, and increased the H2O2 level. Collectively, we efficiently expressed a core peptide of HrpZ and elucidated its strong ability to elicit a HR and resistance to downy mildew. This research provides insight into understanding the structure and function of HrpZ and will advance the application of HrpZ_C_2_2 to increase the resistance of grapevine to downy mildew. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is one of the infectious diseases caused by the pathogen Mycobacterium tuberculosis that continuously threatens the global human health. Bacillus Calmette-Guérin (BCG) vaccine is the only vaccine that has been used clinically to prevent tuberculosis in recent centuries, but its limitations in preventing latent infection and reactivation of tuberculosis do not provide full protection. In this study, we selected the membrane-associated antigen Rv1513 of Mycobacterium. In order to achieve stable expression and function of the target gene, the prokaryotic expression recombinant vector pET30b-Rv1513 was constructed and expressed and purified its protein. Detection of IFN- γ levels in the peripheral blood of TB patients stimulated by whole blood interferon release assay (WBIA) and multi-microsphere flow immunofluorescence luminescence (MFCIA) revealed that the induced production of cytokines, such as IFN-γ and IL-6, was significantly higher than that in the healthy group. Rv1513 combined with adjuvant DMT (adjuvant system liposomes containing dimethyldioctadecylammonium bromide (DDA), monophospholipid A (MPL), and trehalose-660-dibenzoic acid (TDB)) was used to detect serum specific antibodies, cytokine secretion from splenic suprasplenic cell supernatants, and multifunctional T-cell levels in splenocytes in immunised mice. The levels of IFN-γ, TNF-α, and IL-2 secreted by mouse splenocytes were found in the Rv1513+DMT group and the BCG+Rv1513+DMT group. The serum levels of IgG and its subclasses and the number of IFN-γ+T cells, TNF-α+T and IFN-γ+TNF-α+T cells in the induced CD4+/CD8+T cells in mice were significantly higher than those in the BCG group, and the highest levels were found in the BCG+Rv1513+DMT group. These findings suggest that Rv1513/DMT may serve as a potential subunit vaccine candidate that may be effective as a booster vaccine after the first BCG vaccination. [ABSTRACT FROM AUTHOR]

Pinctada fucata meat is the main by-product of the pearl harvesting industry. It is rich in nutrition, containing a lot of protein and peptides, and holds significant value for both medicine and food. In this study, a new active protein was discovered and expressed heterogeneously through bioinformatics analysis. It was then identified using Western blot, molecular weight, and mass spectrometry. The antibacterial activity, hemolysis activity, antioxidant activity, and Angiotensin-Converting Enzyme II (ACE2) inhibitory activity were investigated. An unknown functional protein was screened through the Uniprot protein database, and its primary structure did not resemble existing proteins. It was an α-helical cationic polypeptide we named PFAP-1. The codon-optimized full-length PFAP-1 gene was synthesized and inserted into the prokaryotic expression vector pET-30a. The induced expression conditions were determined with a final isopropyl-β-d-thiogalactoside (IPTG) concentration of 0.2 mM, an induction temperature of 15 °C, and an induction time of 16 h. The recombinant PFAP-1 protein, with low endotoxin and sterility, was successfully prepared. The recombinant PFAP-1 protein exhibited strong antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) in vitro, and the diameter of the inhibition zone was 15.99 ± 0.02 mm. Its minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were 37.5 μg/mL and 150 μg/mL, respectively, and its hemolytic activity was low (11.21%) at the bactericidal concentration. The recombinant PFAP-1 protein significantly inhibited the formation of MRSA biofilm and eradicated MRSA biofilm. It also demonstrated potent 1,1-diphenyl-2-picryl-hydrazyl radical (DPPH) scavenging activity with a half-maximal inhibitory concentration (IC50) of 40.83 μg/mL. The IC50 of ACE2 inhibition was 5.66 μg/mL. Molecular docking results revealed that the optimal docking fraction of PFAP-1 protein and ACE2 protein was −267.78 kcal/mol, with a confidence level of 0.913. The stable binding complex was primarily formed through nine groups of hydrogen bonds, three groups of salt bridges, and numerous hydrophobic interactions. In conclusion, recombinant PFAP-1 can serve as a promising active protein in food, cosmetics, or medicine. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Hydrobiology is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To explore substitutes for natural tropomyosin as basic materials for the diagnosis and treatment of shrimp allergy, total RNA was extracted from Macrobrachium nipponense, and the full-length sequence of the tropomyosin gene was cloned by rapid amplification of cDNA ends (RACE). Specific primers were designed based on the cDNA sequence of tropomyosin and the expression plasmid pET-30a-TM was constructed. Finally, recombinant tropomyosin was expressed under the induction of isopropyl-β-D-thiogalactopyranoside (IPTG). The results showed that the full-length cDNA sequence of tropomyosin was 1 658 bp in length (GenBank accession number: OP974621). Its open reading frame (ORF) was 855 bp in length, encoding 284 amino acids, with a predicted protein isoelectric point of 4.70 and a predicted molecular mass of 32.8 kDa. The highest expression was obtained after 4 h of induction with a final IPTG concentration of 1 mmol/L at 37 ℃. The recombinant tropomyosin mainly existed as a soluble form in the supernatant of cell lysate with a molecular mass of approximately 38 kDa.

Abstract Background The neuroimmune network plays a crucial role in regulating mucosal immune homeostasis within the digestive tract. Synaptosome-associated protein 25 (SNAP-25) is a presynaptic membrane-binding protein that activates ILC2s, initiating the host's anti-parasitic immune response. Methods To investigate the effect of Moniezia benedeni (M. benedeni) infection on the distribution of SNAP-25 in the sheep's small intestine, the recombinant plasmid pET-28a-SNAP-25 was constructed and expressed in BL21, yielding the recombinant protein. Then, the rabbit anti-sheep SNAP-25 polyclonal antibody was prepared and immunofluorescence staining was performed with it. The expression levels of SNAP-25 in the intestines of normal and M. benedeni-infected sheep were detected by ELISA. Results The results showed that the SNAP-25 recombinant protein was 29.3 KDa, the titer of the prepared immune serum reached 1:128,000. It was demonstrated that the rabbit anti-sheep SNAP-25 polyclonal antibody could bind to the natural protein of sheep SNAP-25 specifically. The expression levels of SNAP-25 in the sheep's small intestine revealed its primary presence in the muscular layer and lamina propria, particularly around nerve fibers surrounding the intestinal glands. Average expression levels in the duodenum, jejunum, and ileum were 130.32 pg/mg, 185.71 pg/mg, and 172.68 pg/mg, respectively. Under conditions of M. benedeni infection, the spatial distribution of SNAP-25-expressing nerve fibers remained consistent, but its expression level in each intestine segment was increased significantly (P

[Objective] This study is to identify and analyze the flavonol synthetase FLS gene family as a key enzyme in the secondary metabolism of polyphenols in Caryophyllum plants and to explore the function of FLSs in Chenopodium quinoa. [Method] Bioinformatics analysis website was used to identify the CqFLS family members, and analyze their gene structure, protein physicochemical properties, secondary structure, tertiary structure, promoter cis element and phylogenetic relationship. The gene cloning and expression vector construction was to analyze their protein expression. [Result] The 3 CqFLSs genes were identified, and they were unevenly distributed on two chromosomes, and the promoter region of CqFLSs contained salicylic acid, abscisic acid, methyl jasmonate and drought-induced elements. There were multiple InDel and SNP variations in CqFLS2.1g,the coding nucleotides were deleted at ch01 28871893, 28871125, and 28872881, and all of them were annotated as upstream effects, and no frameshift mutations were detected. Phylogenetic tree analysis showed that CqFLS2.1g were in different branches from CqFLS1.1g and CqFLS3.10g,their expressions varied, and CqFLS2.1g were also differentiated. Meanwhile gene expression analysis revealed that the overall expressions of the three CqFLSs genes in“Qingbai 1”grains were higher than those of“Qinghei 1”and “Gongzha 4”. The cloned CqFLS1.1g was induced with 0.3 mmol/L IPTG and was successfully expressed at both 20℃ and 37℃ . [Conclusion] CqFLS1.1g plays a role in the formation of flowers and grain development, while CqFLS2.1g is mainly involved in the formation of quinoa grains, and the expression of CqFLS is tissue-specific and plays an important role in the growth and development of quinoa [ABSTRACT FROM AUTHOR]

Since 2017, an infectious goose gout disease characterized by urate precipitation in viscera, mainly caused by novel goose astrovirus (GoAstV) infection, has emerged in the main goose-producing region of China. The current challenge in managing goose gout disease is largely due to the absence of a rapid and efficient detection method for the GoAstV pathogen. Notably, the potential application of immunosensors in detecting GoAstV has not yet been explored. Herein, a label-free PEC immunosensor was fabricated by using purchased TiO2 as the photoactive material and antibody against GoAstV P2 proteins as the specific recognition element. First, we successfully expressed the capsid spike domain P2 protein of ORF2 from GoAstV CHSH01 by using the pET prokaryotic expression system. Meanwhile, the polyclonal antibody against GoAstV capsid P2 protein was produced by purified protein. To our knowledge, this is the first establishment and preliminary application of the label-free photoelectrochemical immunosensor method in the detection of AstV. The PEC immunosensor had a linear range of 1.83 fg mL−1 to 3.02 ng mL−1, and the limit of detection (LOD) was as low as 0.61 fg mL−1. This immunosensor exhibited high sensitivity, great specificity, and good stability in detecting GoAstV P2 proteins. To evaluate the practical application of the immunosensor in real-world sample detection, allantoic fluid from goose embryos was collected as test samples. The results indicated that of the eight positive samples, one false negative result was detected, while both negative samples were accurately detected, suggesting that the constructed PEC immunosensor had good applicability and practical application value, providing a platform for the qualitative detection of GoAstV. [ABSTRACT FROM AUTHOR]

Background: The neuroimmune network plays a crucial role in regulating mucosal immune homeostasis within the digestive tract. Synaptosome-associated protein 25 (SNAP-25) is a presynaptic membrane-binding protein that activates ILC2s, initiating the host's anti-parasitic immune response. Methods: To investigate the effect of Moniezia benedeni (M. benedeni) infection on the distribution of SNAP-25 in the sheep's small intestine, the recombinant plasmid pET-28a-SNAP-25 was constructed and expressed in BL21, yielding the recombinant protein. Then, the rabbit anti-sheep SNAP-25 polyclonal antibody was prepared and immunofluorescence staining was performed with it. The expression levels of SNAP-25 in the intestines of normal and M. benedeni-infected sheep were detected by ELISA. Results: The results showed that the SNAP-25 recombinant protein was 29.3 KDa, the titer of the prepared immune serum reached 1:128,000. It was demonstrated that the rabbit anti-sheep SNAP-25 polyclonal antibody could bind to the natural protein of sheep SNAP-25 specifically. The expression levels of SNAP-25 in the sheep's small intestine revealed its primary presence in the muscular layer and lamina propria, particularly around nerve fibers surrounding the intestinal glands. Average expression levels in the duodenum, jejunum, and ileum were 130.32 pg/mg, 185.71 pg/mg, and 172.68 pg/mg, respectively. Under conditions of M. benedeni infection, the spatial distribution of SNAP-25-expressing nerve fibers remained consistent, but its expression level in each intestine segment was increased significantly (P < 0.05), up to 262.02 pg/mg, 276.84 pg/mg, and 326.65 pg/mg in the duodenum, jejunum, and ileum, and it was increased by 101.06%, 49.07%, and 89.16% respectively. Conclusions: These findings suggest that M. benedeni could induce the SNAP-25 expression levels in sheep's intestinal nerves significantly. The results lay a foundation for further exploration of the molecular mechanism by which the gastrointestinal nerve-mucosal immune network perceives parasites in sheep. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The aim of this investigation was to evaluate the antifungal activity of Osmotin against several important economically improtant plant pathogenic fungi. The Osmotin gene from Nicotiana tabacum was overexpressed in Escherichia coli (Rosetta DE3). Taguchi test was applied to optimize the conditions for protein expression, and Western blot analysis confirmed expression of the recombinant protein. The expressed Osmotin was found in the form of insoluble inclusion bodies, which were then solubilized and refolded. The purified and refolded protein's activity was verified by three different antifungal activity assays. The purified recombinant Osmotin demonstrated a wide range of antifungal activity against various types of phytopathogenic fungi: Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium oxysporum, Verticillium dahliae and Alternaria brassicola. Hemolysis biosafety test demonstrated that the protein is non-toxic to mammalian cells. All these findings suggest that the Osmotin gene from N. tabacum has promising potential as an antifungal agent against various phytopathogenic fungi and also to be utilized for production of antifungal agents and fungal-resistant transgenic plants. [ABSTRACT FROM AUTHOR]

Disintegrins, a family of snake venom protein, which are capable of modulating the activity of integrins that play a fundamental role in the regulation of many physiological and pathological processes. The main purpose of this study is to obtain the recombinant disintegrin (r-DI) and evaluate its biological activity. In this study, we explored a high-level expression prokaryotic system and purification strategy for r-DI. Then, r-DI was treated to assay effects on cell growth, migration, and invasion. The affinity for the interactions of r-DI with integrin was determined using Surface plasmon resonance (SPR) analyses. The r-DI can be expressed in Escherichia coli and purified by one-step chromatography. The r-DI can inhibit B16F10 cells proliferation, migration, and invasion. Also, we found that r-DI could interact with the integrin αIIbβ3 (GPIIb/IIIa). The r-DI can be expressed, purified, characterized through functional assays, and can also maintain strong biological activities. Thus, this study showed potential therapeutic effects of r-DI for further functional and structural studies. [ABSTRACT FROM AUTHOR]

[Objective] The expression pattern and protein characteristics of the B-galactosidase gene NIBGAL from Nicotiana tabacum were explored to provide a basis for the future development of tobacco as a versatile platform material. (Method) Through comparing the homologous between the gene sequences of N. tabacum K326 and NbBGALI gene of N. benthamiana, NIBGAL gene was cloned through PCR by taking cDNA of K326 as the template. Subsequently, bioinformatics tools were used to analyze its gene structure and protein physicochemical properties. The gene expression pattern in different tobacco tissues (roots, stems, leaves and flowers) was examined using qRT-PCR. Finally, the research on prokaryotic expression, protein purification and protein characterization of the gene was carried out. (Result) The cloned tobacco NIBGAL gene obtained in the study exhibited the highest homology with the NbBGALI gene in N. benthamiana, and both possessed identical structural domains (Motif). The gene was located on chromosome 9, and it contained 18 introns. The mRNA length of the gene was 6649 bp. The CDS was 2541 bp encoding 846 amino acids. The enzyme belonged to the GH35 family. The NtBGAL protein had a molecular weight of 92.44 kDa, a theoretical isoelectric point (pl) of 6.8, GRAVY score of -0.222, and was localized on the cell wall. There was a significant difference in gene expression levels of NIBGAL in different tissues of N. tabacum, followed by flower > leaf > stem > root. The prokaryotic expression was performed at 37 °C, NtBGAL protein had obvious expression, and the higher purified NIBGAL protein was obtained by purification method of inclusion body. The optimal temperature for the enzymatic reaction of NtBGAL protein was 30-40 °C and the optimal pH was 4.0-6.5. NIBGAL enzyme activity was enhanced at Mn2+ concentrations ranging from 0.05 to 0.15 mmol/L. [Conclusion] The study clarifies the tissue expression pattern and protein characterization of the NIBGAL gene, which lays the foundation for further modification of the NIBGAL gene to produce humanized glycoproteins in N. tabacum. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Shenyang Agricultural University is the property of Journal of Shenyang Agricultural University Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

[Objective]This work aims to explore the role of chalcone synthase(chalcone synthase，PcCHS)gene in the synthesis of flavonoids in Polygonatum cyrtonema Hua, which may provide a reliable theoretical basis for the subsequent analysis of the function of PcCHS and the breeding of new varieties of P. cyrtonema Hua.[Method]Using P. cyrtonema Hua as cDNA template, the coding sequence of PcCHS gene was cloned, and the gene was analyzed bioinformatically. The prokaryotic expression vector of PcCHS was constructed and the recombinant protein was purified to verify the expression activity of PcCHS in vitro. Transient overexpression system was used to investigate the changes of total flavonoids content after overexpression of this gene. Gateway technology was used to construct the subcellular localization vector 35S::PcCHS-GFP, and the subcellular localization of target protein was determined by the tobacco expression system.[Result]The results showed that PcCHS was a hydrophilic protein with an open reading frame of 1 251 bp, a theoretical molecular weight of 44.63 kD and an isoelectric point of 5.89, and was closely related to AoCHS(Asparagus officinalis). Prokaryotic expression experiment showed that pET28aPcCHS was induced to express soluble recombinant protein by IPTG(isopropyl-β-d-thiogalactoside). Western-blot showed that the size of pET28a-PcCHS was about 45 kD, which was consistent with the expected size. The purified protein had certain enzymatic activity and catalyzed the conversion of p-coumaryl-CoA and malonyl-CoA into naringin chalcone. In addition, in the transient overexpression of PcCHS, the expression level of PcCHS group was significantly higher than that of no-load K group, and the total flavonol content was also significantly higher than that of no-load K group, up to 1.83 times. Subcellular localization results showed that the gene plays a role in the cell membrane and nucleus. [Conclusion]Prokaryotic expression of PcCHS gene has enzyme activity in vitro, and its subcellular location is in cell membrane and nucleus, and instantaneous overexpression significantly increase the total flavonoid content in the leaves of P. cyrtonema Hua. [ABSTRACT FROM AUTHOR]

[Objective]The objective of this study is to identify the genes(TaENO2s)encoding the enolase family member ENO2 in the important crop plant of wheat(Triticum aestivum L.)and to determine the alternative translation feature and protein enolase activity of TaENO2s.[Method]Wheat TaENO2s were identified by bioinformatics. One of the identified TaENO2s was selected as a representative for protoplast-based exogenous expression and characterization of alternative translation products. This selected TaENO2 was also subjected to prokaryotic expression for purification of recombinant protein. The enolase activity of the purified recombinant protein was determined with in vitro assays.[Result]The hexaploid wheat genome had three homeologous TaENO2s which sequences encoded a conserved enolase catalytic center and had a putative alternative translation start site at the internal ATG codon encoding the residue M94(ATGM94) of the enolase sequence. When exogenously expressed in protoplasts, the representative TaENO2 encoded two proteins, one full-length form of cytoplasmic and nuclear enolase protein and one N-terminal truncated form of nuclear transcriptional repressor TaMBP-1, while the variant of the same gene containing mutated ATGM94 only encoded the full-length form of enolase protein. Soluble recombinant GST-TaENO2 protein expressed in Escherichia coli was successfully purified and verified to have an in vitro enzymatic activity to catalyze the conversion from 2-phosphoglycerate (2-PGA)to phosphoenolpyruvate(PEP)[Conclusion]The wheat genome has three TaENO2 genes encoding the enolase protein ENO2. Under the condition of exogenous expression, TaENO2 encodes two protein products via mRNA alternative translation, one enolase protein translated from the first start codon and one TaMBP-1 protein translated from the internal start codon(2-PGA)to phosphoenolpyruvate(PEP).[Conclusion[The wheat genome has three TaENO2 genes encoding the enolase protein ENO2. Under the condition of exogenous expression, TaENO2 encodes two protein products via mRNA alternative translation, one enolase protein translated from the first start codon and one TaMBP-1 protein translated from the internal start codon ATGM94. The prokaryotically expressed recombinant protein GST-TaENO2 possesses in vitro enolase activity.. The prokaryotically expressed recombinant protein GST-TaENO2 possesses in vitro enolase activity. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao is the property of Chinese Journal of Preventive Veterinary Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Food Science & Biotechnology is the property of Journal of Food Science & Biotechnology Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

【Objective】This study aims to clone the sterol glycosyltransferase gene AoSGT1 from Asparagus officinalis, to assess its catalytic properties, and to elucidate its role in the biosynthesis of asparagus saponins and their metabolic regulation.【Method】We designed specific primers based on asparagus transcriptome data to amplify the complete open reading frame（ORF）of AoSGT1. The gene was sequenced and analyzed through bioinformatics, and its expression profile was investigated via quantitative real-time PCR（RT-qPCR）. We also constructed a pGEX-4T-3-AoSGT1 expression vector, expressed it in Escherichia coli BL21（DE3）, and induced the recombinant protein production.【Result】AoSGT1 has a length of 1 800 base pairs, encoding 599 amino acids, with a relative molecular weight of 66.72 kD. It is a hydrophilic protein with no transmembrane domains or signal peptides. The phylogenetic analysis revealed a close homology between AoSGT1 and Dioscorea zingiberensis Dz3GT2, indicating their membership in the UGT80B1 subfamily. Multiple sequence alignment revealed that the protein sequence contained conserved domains“PSBD Box”and“PSPG Box”of sterol glycosyltransferase, indicating its potential glycosylation activity at the 3β-OH position of steroidal compounds. RT-qPCR results showed that AoSGT1 was highly expressed in the roots, while expression in stems and flowers was relatively low. Additionally, SDS-PAGE results revealed that the target protein was efficiently expressed in a soluble form within E. coli, matching the predicted size.【Conclusion】The AoSGT1 gene was successfully cloned and demonstrated to exhibit tissue-specific expression in asparagus, suggesting its potential involvement in the biosynthesis of steroidal saponins in asparagus. Additionally, heterologous expression of the target protein in E. coli was successfully achieved. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Northwest A & F University - Natural Science Edition is the property of Editorial Department of Journal of Northwest A&F University (Natural Science Edition) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Context. The cDNA sequence of hepatocyte growth factor (HGF) gene in Tarim red deer has not been reported yet. Aims. This study aims to obtain the full-length cDNA sequence of HGF and analyse its expression in different parts of developing antler tissues. The result provides foundational data for understanding the potential role of HGF in regulating antler growth and development. Methods. Rapid amplification of cDNA ends was used to obtain the full-length cDNA sequence of Tarim red deer HGF. The pET28a (+) vector was constructed for prokaryotic expression of the recombinant protein in E. coli BL21 (DE3). The expression of HGF in different antler tissues was detected using real-time quantitative polymerase chain reaction and immunohistochemistry. Key results. The full-length cDNA of Tarim red deer HGF was found to consist of a 156 bp 5'untranslated region (UTR), a 112 bp 3'UTR, and a 2193 bp open reading frame encoding a protein of 730 amino acids. The recombinant HGF protein expressed in prokaryotic cells formed inclusion bodies. HGF and its receptor c-Met were expressed in all four different antler tissues, with the highest expression level in velvet skin, followed by bone, cartilage, and the lowest in the mesenchyme. Conclusions. The study successfully obtained the full-length cDNA sequence of Tarim red deer HGF and identified the expression profile of HGF and c-Met in different antler tissues. HGF is a candidate gene that may play a role in regulating the growth and development of deer antler. Implications. These findings provide fundamental data for further investigations into the role of HGF in antler development. Understanding the function of HGF in antler development could have implications for elucidating the mechanism of antler regeneration. [ABSTRACT FROM AUTHOR]

Objective To express the protein enconded by the Rv3432c gene of Mycobacterium tuberculosis (M.tb) in vitro by prokaryotic expression, to analyze the structure of the Rv3432c protein by using bioinformatics software, and to explore for new drug targets against M.tb. Methods The Rv3432c gene was amplified by PCR using the genomic DNA of the inactivated M.tb strain H37Rv as the template and a recombinant plasmid was constructed with the expression vector pET-28a. The expression products were analyzed by SDS-PAGE and purified using affinity chromatography. The biological properties of Rv3432c were analyzed with Protparam, the Pfam online tool, SOMPA, Protscale, TMHMM Signalp 6.0, NetPhos3.1, SUMOsp 2.0, and SWISS-MODEL. Results pET-28a-Rv3432c recombinant plasmid sequencing results were fully consistent with those of the target gene. SDS-PAGE analysis showed that the fusion protein existed in the form of a soluble protein with a relative molecular mass of about 55×10³, which matched the expected size. ProtParam analysis showed that the Rv3432c protein was hydrophilic (showing a GRAVY value of -0.079). Rv3432c was a protein with no transmembrane structural domains or signal peptide. The secondary structure of Rv3432c mainly consisted of random coils (39.78%) and α-helix (39.57%) and was relatively loosely structured. Conclusion We successfully constructed a prokaryotic expression plasmid of the Rv3432c protein and analyzed its structure using bioinformatics, laying the foundation for further research on the role of Rv3432c in the pathogenesis and progression of tuberculosis as well as the identification of new drug targets against M.tb. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Northwest A & F University - Natural Science Edition is the property of Editorial Department of Journal of Northwest A&F University (Natural Science Edition) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao is the property of Chinese Journal of Preventive Veterinary Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Hepatitis-hydropericardium syndrome (HHS), caused by fowl adenovirus serotype 4 (FAdV-4), has been widely spread across China, resulting in great financial losses in the poultry industry. Therefore, efficient vaccines against this disease urgently need to be developed. In our study, the fiber-2 and penton base proteins derived from the FAdV-4 JS strain were expressed in a prokaryotic system (E. coli) in a soluble form. Then, the efficacy of the two recombinant proteins formulated with cheap and widely used adjuvants (Marcol™ 52 white oil) were respectively tested, and the minimum immune doses and safety of the above proteins were also determined. It was indicated that the fiber-2 (20 µg/bird, 200 µg/bird) and penton base (200 µg/bird) could provide complete protection against the highly pathogenic FAdV-4 and suppress its replication and shedding. Unfortunately, only the fiber-2 protein could induce complete protection (10/10) at a low dose (10 µg/bird). In addition, we confirmed that the fiber-2 subunit vaccine formulated with oil adjuvants was safe for vaccinated chickens. Conclusively, all of our results suggest that we successfully prepared an efficient and cheap fiber-2 subunit vaccine with few side effects. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Food Science & Biotechnology is the property of Journal of Food Science & Biotechnology Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

SUMMARY: Avian influenza viruses (AIV) of the H3N8 subtype pose a significant threat to both the poultry industry and public health. This study aimed to develop and validate a highly specific polyclonal antibody targeting the neuraminidase (NA) protein from a novel H3N8 AIV, which exhibits tri-basic hemagglutinin cleavage sites and shares genetic proximity to recent human isolates. The NA gene of H3N8 AIV was cloned and introduced into E. coli BL21 and Rosetta competent cells to induce the recombinant protein expression. Optimization procedures, including IPTG concentration, time, and temperature, were implemented to enhance protein expression efficiency. Polyclonal antibodies were generated and validated through western blotting, indirect immunofluorescence assay (IFA), and indirect ELISA. As a result, the pET-32a-NA (N8) vector was successfully constructed. The expression of recombinant NA protein with a size of approximately 70 KDa was obtained and then optimized with a final IPTG concentration of 0.6 mM, at 27°C for 14 h. Western blotting and IFA analysis demonstrated that the prepared polyclonal antibody effectively and specifically bound to NA(N8) protein. The titer of the polyclonal antibodies reached 1:409600 by indirect ELISA. These results indicate the potential of these antibodies for the development of detection assays and biological studies required for H3N8 AIVs.

Abstract Background Gelsemium elegans is a traditional Chinese medicinal plant and temperature is one of the key factors affecting its growth. RAV (related to ABI3/VP1) transcription factor plays multiple roles in higher plants, including the regulation of plant growth, development, and stress response. However, RAV transcription factor in G. elegans has not been reported. Results In this study, three novel GeRAV genes (GeRAV1-GeRAV3) were identified from the transcriptome of G. elegans under low temperature stress. Phylogenetic analysis showed that GeRAV1-GeRAV3 proteins were clustered into groups II, IV, and V, respectively. RNA-sequencing (RNA-seq) and real-time quantitative PCR (qRT-PCR) analyses indicated that the expression of GeRAV1 and GeRAV2 was increased in response to cold stress. Furthermore, the GeRAV1 gene was successfully cloned from G. elegans leaf. It encoded a hydrophilic, unstable, and non-secretory protein that contained both AP2 and B3 domains. The amino acid sequence of GeRAV1 protein shared a high similarity of 81.97% with Camptotheca acuminata CaRAV. Subcellular localization and transcriptional self-activation experiments demonstrated that GeRAV1 was a nucleoprotein without self-activating activity. The GeRAV1 gene was constitutively expressed in the leaves, stems, and roots of the G. elegans, with the highest expression levels in roots. In addition, the expression of the GeRAV1 gene was rapidly up-regulated under abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (MeJA) stresses, suggesting that it may be involved in hormonal signaling pathways. Moreover, GeRAV1 conferred improved cold and sodium chloride tolerance in Escherichia coli Rosetta cells. Conclusions These findings provided a foundation for further understanding on the function and regulatory mechanism of the GeRAV1 gene in response to low-temperature stress in G. elegans.

Abstract Background Glutathione S-transferases (GSTs) are a superfamily of multifunctional enzymes in living organisms with metabolic and detoxification functions, which can detoxify exogenous and endogenous compounds and thereby reduce the damage caused by toxic substances to the body. Ticks are obligate blood-sucking ectoparasites that can transmit various pathogens, and the characterization of tick-derived GSTs may help improve current understanding of the molecular mechanism of tick resistance to insecticides. In this study, a novel GST gene, named HrGSTm1, was identified from Hyalomma rufipes. Methods Sequence analysis was performed by using bioinformatics techniques. A prokaryotic expression system was used to obtain the recombinant expression protein rHrGSTm1. Detection of spatiotemporal expression patterns of target genes and their response to the toxicity of cyhalothrin on female H. rufipes was performed by using a quantitative PCR platform. The optimal enzymological parameters of rHrGSTm1 using glutathione as substrate were calculated. The antioxidant capacity of the recombinant protein was evaluated by DPPH• (1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl). Knockdown of the HrGSTm1 genes through RNA interference was used to analyze their effects on the physiological parameters of ticks. The changes in HrGSTm1 messenger RNA expression patterns under cypermethrin stress were analyzed. Results The complementary DNA sequence of HrGSTm1 contained a 672-bp open reading frame, which potentially encoded 223 amino acids. The predicted molecular weight was 25.62 kDa, and the isoelectric point 8.22. HrGSTm1 is a Mu-class GST, belonging to the cytoplasmic GSTs with no signal peptide observed. The V max and K m of rHrGSTm1 were 3.367 ± 0.81 uM and 2.208 ± 0.76 uM, respectively, and its activities were dependent on different temperatures and pH conditions; the scavenging rate of rHrGSTm1 to DPPH• reached 76.4% at 1.25 mg/ml. Variable expressions of HrGSTm1 were observed under various treatment periods and in different tissues, with the highest appearing in eggs (analysis of variance [ANOVA], F (2, 9) = 279.9, P

Objective: To express the protein HmpA of Salmonella paratyphi A in prokaryotes, and perform bioinformatics analysis on it to provide a theoretical reference for studying the effect of this protein on the nitric oxide (NO) signaling pathway in the host. Methods: The hmpA gene was amplified by PCR and subcloned into the T-vector. Then the expression vector pNdeI-hmpA was constructed and transformed into BL21 (DE3). After induced by IPTG, the recombinant protein expression form was assessed by SDS-PAGE. HmpA was purified using Histrap preload column and identified by Western blot. Bioinformatics technology was used to analyze the characteristics of HmpA. Results: The prokaryotic expression vector pNdeI-hmpA was successfully constructed. HmpA coexisted in the inclusion body and soluble forms under low temperatures after being induced with IPTG. The purified HmpA protein could be detected by the anti-His antibody. Bioinformatics analysis suggested that HmpA was a hydrophilic protein with no transmembrane structural domain or signal peptide. The secondary structure of this protein was mainly composed of α-helix with irregular convolutions, and the tertiary structure model was similar to a ring. The protein structural domain analysis showed that HmpA contains one functional structural domain belonging to the PRK13289 superfamily. There were 32 phosphorylation sites in HmpA. Associated with multiple Salmonella proteases or transcription factors that degrade nitrates and killer NO. Conclusion: This study successfully obtained Salmonella paratyphi A HmpA protein by genetic engineering technology and predicted part of its biological characteristics by bioinformatics methods, which would provide theoretical support for the subsequent uncovering of the influence of HmpA of Salmonella paratyphi A on the nitric oxide signaling pathway in the host.

Cysteine plays a pivotal role in the sulfur metabolism network of plants, intimately influencing the conversion rate of organic sulfur and the plant’s capacity to withstand abiotic stresses. In tea plants, the serine acetyltransferase (SAT) genes emerge as a crucial regulator of cysteine metabolism, albeit with a notable lack of comprehensive research. Utilizing Hidden Markov Models, we identified seven CssSATs genes within the tea plant genome. The results of the bioinformatics analysis indicate that these genes exhibit an average molecular weight of 33.22 kD and cluster into three distinct groups. Regarding gene structure, CssSAT1 stands out with ten exons, significantly more than its family members. In the promoter regions, cis-acting elements associated with environmental responsiveness and hormone induction predominate, accounting for 34.4% and 53.1%, respectively. Transcriptome data revealed intricate expression dynamics of CssSATs under various stress conditions (e.g., PEG, NaCl, Cold, MeJA) and their tissue-specific expression patterns in tea plants. Notably, qRT-PCR analysis indicated that under salt stress, CssSAT1 and CssSAT3 expression levels markedly increased, whereas CssSAT2 displayed a downregulatory trend. Furthermore, we cloned CssSAT1-CssSAT3 genes and constructed corresponding prokaryotic expression vectors. The resultant recombinant proteins, upon induction, significantly enhanced the NaCl tolerance of Escherichia coli BL21, suggesting the potential application of CssSATs in bolstering plant stress resistance. These findings have enriched our comprehension of the multifaceted roles played by CssSATs genes in stress tolerance mechanisms, laying a theoretical groundwork for future scientific endeavors and research pursuits.

Background: Gelsemium elegans is a traditional Chinese medicinal plant and temperature is one of the key factors affecting its growth. RAV (related to ABI3/VP1) transcription factor plays multiple roles in higher plants, including the regulation of plant growth, development, and stress response. However, RAV transcription factor in G. elegans has not been reported. Results: In this study, three novel GeRAV genes (GeRAV1-GeRAV3) were identified from the transcriptome of G. elegans under low temperature stress. Phylogenetic analysis showed that GeRAV1-GeRAV3 proteins were clustered into groups II, IV, and V, respectively. RNA-sequencing (RNA-seq) and real-time quantitative PCR (qRT-PCR) analyses indicated that the expression of GeRAV1 and GeRAV2 was increased in response to cold stress. Furthermore, the GeRAV1 gene was successfully cloned from G. elegans leaf. It encoded a hydrophilic, unstable, and non-secretory protein that contained both AP2 and B3 domains. The amino acid sequence of GeRAV1 protein shared a high similarity of 81.97% with Camptotheca acuminata CaRAV. Subcellular localization and transcriptional self-activation experiments demonstrated that GeRAV1 was a nucleoprotein without self-activating activity. The GeRAV1 gene was constitutively expressed in the leaves, stems, and roots of the G. elegans, with the highest expression levels in roots. In addition, the expression of the GeRAV1 gene was rapidly up-regulated under abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (MeJA) stresses, suggesting that it may be involved in hormonal signaling pathways. Moreover, GeRAV1 conferred improved cold and sodium chloride tolerance in Escherichia coli Rosetta cells. Conclusions: These findings provided a foundation for further understanding on the function and regulatory mechanism of the GeRAV1 gene in response to low-temperature stress in G. elegans. [ABSTRACT FROM AUTHOR]

Background: Glutathione S-transferases (GSTs) are a superfamily of multifunctional enzymes in living organisms with metabolic and detoxification functions, which can detoxify exogenous and endogenous compounds and thereby reduce the damage caused by toxic substances to the body. Ticks are obligate blood-sucking ectoparasites that can transmit various pathogens, and the characterization of tick-derived GSTs may help improve current understanding of the molecular mechanism of tick resistance to insecticides. In this study, a novel GST gene, named HrGSTm1, was identified from Hyalomma rufipes. Methods: Sequence analysis was performed by using bioinformatics techniques. A prokaryotic expression system was used to obtain the recombinant expression protein rHrGSTm1. Detection of spatiotemporal expression patterns of target genes and their response to the toxicity of cyhalothrin on female H. rufipes was performed by using a quantitative PCR platform. The optimal enzymological parameters of rHrGSTm1 using glutathione as substrate were calculated. The antioxidant capacity of the recombinant protein was evaluated by DPPH• (1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl). Knockdown of the HrGSTm1 genes through RNA interference was used to analyze their effects on the physiological parameters of ticks. The changes in HrGSTm1 messenger RNA expression patterns under cypermethrin stress were analyzed. Results: The complementary DNA sequence of HrGSTm1 contained a 672-bp open reading frame, which potentially encoded 223 amino acids. The predicted molecular weight was 25.62 kDa, and the isoelectric point 8.22. HrGSTm1 is a Mu-class GST, belonging to the cytoplasmic GSTs with no signal peptide observed. The Vmax and Km of rHrGSTm1 were 3.367 ± 0.81 uM and 2.208 ± 0.76 uM, respectively, and its activities were dependent on different temperatures and pH conditions; the scavenging rate of rHrGSTm1 to DPPH• reached 76.4% at 1.25 mg/ml. Variable expressions of HrGSTm1 were observed under various treatment periods and in different tissues, with the highest appearing in eggs (analysis of variance [ANOVA], F(2, 9) = 279.9, P < 0.0001) and Malpighian tubules (ANOVA, F(3, 12) = 290.5, P < 0.0001). After knockdown of HrGSTm1, compared with the control group, the mortality in the treatment group was increased by 16.7%, the average oviposition rate decreased by 33.9%, the average engorged body weight decreased by 287.38 mg and egg weight decreased by 127.46 mg, although only the engorged body weight was significantly different (t-test, t(44) = 2.886, P = 0.006). After exposure to three sublethal concentrations (LC05, LC10, LC50) of cyhalothrin, the expression level of HrGSTm1 in the midgut, ovary and salivary gland was upregulated, whereas in Malpighian tubules, it showed a trend of upregulation at first and then downregulation, implying different functions during the detoxification in different tissues. Conclusions: In this study, a novel GST of the Mu-class was successfully isolated from H. rufipes and systematically subjected to bioinformatic analysis and recombination identification. The variation trend of HrGSTm1 expression level in different tissues suggests that the gene has different detoxification functions in different tissues. The potential function of this gene was analyzed to provide basic research for further investigation of its detoxification mechanism. [ABSTRACT FROM AUTHOR]

Erwinia beijingensis can cause the bacterial soft rot disease of Pleurotus eryngii. In order to clarify the function of glycosyltransferase in E. beijingensis, a recombinant expression vector was constructed to express and purify the glycosyltransferase in this pathogen. The protein activity was measured, and the interactions between proteins were analyzed. The results showed that the prokaryotic expression vector was successfully constructed, 4 soluble proteins were obtained, and MshA, WbnH2, EpsH and TuaG proteins were purified by affinity chromatography column. The activity assay showed that four proteins preferentially used UDP galactose. GST pull down results confirmed that WbnH2 and TuaG proteins interacted with MshA and EpsH proteins in vitro, respectively. Above results laid a foundation for the subsequent study on the function of the glycosyltransferase gene in E. beijingensis. [ABSTRACT FROM AUTHOR]

In this study, we recombinantly expressed the V protein of the peste des petits ruminants virus (PPRV) and evaluated its diagnostic value for PPRV infection using an indirect ELISA (i-ELISA). The optimal concentration of the coated antigen of V protein was 15 ng/well at a serum dilution of 1:400, and the optimal positive threshold value was 0.233. A cross-reactivity assay showed that the V protein-based i-ELISA was specific to PPRV with consistent reproducibility and showed a specificity of 82.6% and a sensitivity of 100% with a virus neutralization test. Using the recombinant V protein as an antigen in ELISA is useful for seroepidemiological studies of PPRV infections. [ABSTRACT FROM AUTHOR]

The acetylcholinesterase (AChE) is involved in termination of synaptic transmission at cholinergic synapses and plays a vital role in the insecticide detection and inhibitor screening. Here, we report the heterologous expression of an AChE from Tetronarce californica (TcA) in Escherichia coli (E. coli) as a soluble active protein. TcA was immobilized in calcium alginate beads; the morphology, biochemical properties, and insecticide detection performance of free and immobilized TcA were characterized. Moreover, we used sequence, structure-based approaches, and molecular docking to investigate structural and functional characterization of TcA. The results showed that TcA exhibited a specific activity of 102 U/mg, with optimal activity at pH 8.0 and 30 °C. Immobilized TcA demonstrated superior thermal stability, pH stability, and storage stability compared to the free enzyme. The highest sensitivity of free TcA was observed with trichlorfon, whereas immobilized TcA showed reduced IC50 values towards tested insecticides by 3 to 180-fold. Molecular docking analysis revealed the interaction of trichlorfon, acephate, isoprocarb, λ-cyhalothrin, and fenpropathrin in the active site gorge of TcA, particularly mediated through the formation of hydrogen bonds and π-π stacking. Therefore, TcA expressed heterologously in E. coli is a promising candidate for applications in food safety and environmental analysis. Key points: • T. californica AChE was expressed solubly in prokaryotic system. • The biochemical properties of free/immobilized enzyme were characterized. • The sensitivity of enzyme to insecticides was evaluated in vitro and in silico. [ABSTRACT FROM AUTHOR]

Copyright of Science & Technology of Food Industry is the property of Science & Technology of Food Industry Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Metacaspases (MCs) is a regulatory gene of programmed cell death in plants, and it participates in the response of plants to stress. MaMC6, a member of MaMCs family, was cloned from Brazilian banana (Musa AAA Cavendishcv. Brazil) cells by PCR, and its bioinformatics analysis, expression pattern analysis and functional verification were carried out. The results showed that MaMC6 was composed of 320 amino acids, was a stable protein, hydrophilic, of no transmembrane structure, contained a caspase‑like domain, belonged to type II metacaspase protein, and had a defense and stress response. The results of RT‑qPCR analysis showed that the relative expression of MaMC6 after 100 mmol/L NaCl treatment was 7.70 times higher than that of the control at 2 h, and the gene expression at 2 h after CaCl2 treatment under salt stress was 3.99 times higher than that of the control, while the use of calcium chelating agent EGTA and calcium channel blocker LaCl3 increased the expression of MaMC6 under salt stress, reaching 9.92 and 11.20 times of the control at 2 h. The results of subcellular localization showed that MaMC6 was located in the cytoplasm and nucleus. The results of E. coli transformation showed that the growth of recombinant strain pET28a‑ MaMC6 was better than that of control strain pET28a under NaCl, mannitol, and high‑temperature stress. To sum up, MaMC6 may be involved in the response of banana to abiotic stress. This study provides a reference for follow‑up study on the role of MaMCs in banana stress resistance. [ABSTRACT FROM AUTHOR]

Copyright of China Tropical Medicine is the property of China Tropical Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Edulis Fungi is the property of Acta Edulis Fungi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)