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The molecular diagnosis of facioscapulohumeral muscular dystrophy (FSHD) relies on detecting contractions of the unique D4Z4 repeat array at the chromosome 4q35 locus in the presence of a permissive 4q35A haplotype. Long, intact DNA molecules are required for accurate sizing of D4Z4 repeats. We validated the use of optical genome mapping to determine size and haplotype of D4Z4 alleles for FSHD analysis. The cohort included 36 unique DNA specimens from fresh blood samples or archived agarose plugs. High-molecular- weight DNA underwent sequence-specific labeling followed by separation and image analysis with data collection on the Saphyr system. D4Z4 allele sizes were calculated and haplotypes determined from the labeling patterns. Each specimen had previous diagnostic testing using restriction enzyme digests with EcoRI, EcoRI/BlnI, XapI, or HindIII, followed by pulsed field gel electrophoresis and Southern blot analysis with appropriate probes. Optical genome mapping detected 4q35 and 10q26 alleles ranging from 1 to 79 D4Z4 repeats and showed strong correlation with Southern blot allele sizing (R(2) = 0.95) and haplotyping (133 of 134; 99.4% haplotype match). Analysis of inter-assay and intra-assay runs showed high reproducibility (0.03 to 0.94 %CV). Subsequent optical genome mapping for routine clinical testing from 315 clinical FSHD cases compared favorably with historical result trends. Optical genome mapping is an accurate and highly reproducible method for chromosomal abnormalities associated with FSHD.

The Tg(Adipoq-cre)1Evdr mouse has become an important tool in adipose tissue biology. However, the exact genomic transgene integration site has not been established. Using Targeted Locus Amplification (TLA) we found the transgene had integrated on mouse chromosome 9 between exons 6 and 7 of Tbx18. We detected transgene-transgene fusion; therefore, we used droplet digital polymerase chain reaction to identify Cre copy number. In two separate experiments, we digested with BAMHI and with HindIII to separate potentially conjoined Cre sequences. We found one copy of intact Cre present in each experiment, indicating transgene-transgene fusion in other parts of the BAC that would not contribute to tissue-specific Cre expression. Cre copy number for Tg(Adipoq-cre)1Evdr mice can be potentially used to identify homozygous mice.

Background Various polymerase chain reaction (PCR)-based methods have been applied for the development of genome walking (GW) technique. These methods which could be based on the application of restriction enzymes or primers have various efficiencies to identify the unknown nucleotide sequences. The present study was conducted to design a new innovative double-strand adaptor using MAP30 gene sequence of Momordica charantia plant as a model to improve genome walking with convenient PCR. Results The adaptor was designed using multiple restriction sites of Hind III, BamH I, EcoR I, and Bgl II enzymes with no restriction site in a known sequence of the MAP30 gene. In addition, no modification was required to add phosphate, amine, or other groups to the adaptor, since restriction enzyme digestion of double-strand adaptor provided the 5′ phosphate group. Here, preparation of the phosphate group in the genomic DNA of the plant digestion with restriction enzymes was performed followed by ligation with digested adaptor containing 5′ phosphate group. Conclusion PCR was done to amplify the unknown sequence using MAP30 gene-specific primer and adaptor primer. Results confirmed the ability of the technique for successful identification of the sequence. Consequently, a newly designed adaptor in the developed technique reduced the time and cost of the method compared to the conventional genome walking; also, cloning and culturing of bacterial steps could be eliminated.

The present study was aimed to evaluate the association of lipoprotein lipase (LPL) gene (S447X and Hind III) polymorphisms and T2DM. Relevant studies were identified through systematic search PubMed, Cochrane Library, Embase, Wanfang, CNKI databases. A total of 22 studies (8 studies for LPL S447X and 14 studies for Hind III) were included. The results showed that the LPL S447X polymorphism was associated with the low risk of T2DM under dominant and allelic genetic models. Subgroup analysis by ethnicity showed that the LPL S447X polymorphism was associated with a decreased risk of T2DM in the Asian population (under dominant, heterozygous and allelic genetic models). In addition, we found that X allele carriers of S447X polymorphism is associated with low levels of TC, TG, and LDL. In subgroup analysis, Hind III polymorphism was associated with low risk of T2DM in Asian populations (under dominant, heterozygote, allele genetic models). Moreover, the carriers of H allele of Hind III have lower levels of TG, and higher levels of HDL-C. This meta-analysis demonstrated that 447X carriers and H allele in LPL gene associated with low risk of T2DM, which may due to in part to the change of serum level of TC, TG, LDL, and HDL.

We studied how the interaction between HindIII endonuclease and dsDNA is affected by the single-base modification of the latter by a single-molecule kinetic assay. For a comparative study of chemical modifications, we measured the binding and unbinding rates of the HindIII-DNA complex for normal dsDNA, methylated DNA, and hydroxymethylated DNA. We found that methylation of DNA at the recognition site results in a large increase in the unbinding rate due to the steric effect, which is consistent with the standard free energy change in the transition state. On the contrary, methylation minimally affects the binding rate, as simultaneous increases in the activation energy and the pre-exponential factor compensate for each other.

Introduction: Ischemic stroke is characterized as a sudden neurological deficit attributed to an acute focal injury of the central nervous system by a vascular cause. This study was performed to determine the frequency of G20210A mutation in the prothrombin gene and its effectiveness on the incidence of ischemic stroke in the Erbil city of Kurdistan region, Iraq. Methods: A total of 50 patients with ischemic stroke was analyzed for the detection of prothrombin gene mutation (G20210A), using polymerase chain reaction (PCR), Restriction fragment length polymorphism (RFLP) with Hind III restriction enzyme. Results: We observed no evidence of an association between ischemic stroke and G20210A mutation in the prothrombin gene in this region. Conclusion: Our finding demonstrates that prothrombotic gene variant seems not to be linked to the incidence of ischemic stroke in Erbil region.

Metapenaeus affinis a species from Metapenaeus genera of Penaeidae shrimps in khaur Abdullah (salty water southern of Iraq), morphologically closely related organism with suspected member in Shatt al-Arab(freshwater)also southern of Iraq. Genetic Characterization for them was study to determine the genetically differences by using (RFLP & RAPD)/PCR molecular markers.The genetic heterogeneity at the DNA level was high in the two organisms in spite of Metapenaes genus share considerable external features. In RFLP marker, voucher DAAPV F7 in 16S rRNA gene was used which was amplified then digested by using the enzymes (SmI ,TagI and HindIII). Restriction patterns of TaqI, SmI & Hind III digested voucher DAAPV F7 in 16S rRNA gene, were clearly differentiate both Metapenaeus affinis and suspected one. Size pieces which were cut result in differing size and differing numbers of patterns Whereas a species which suspected to be Metapenaeus affinis morphologically, gave different results. Overlapping patterns between different species not observed. RFLP results were confirmed the RAPD-PCR results where PCR was carried out using standers conditions for 20 random oligonucleotide primers on gDNA samples of two organisms .Out of them, twelve primers produced high reproducibility and consistent with RAPD profile and yielded monomorphic as well as polymorphic fragments ,while 8 primers yielded either weak amplifications with ambiguity or no amplification at both. The present RAPD study had shown a higher level of polymorphism in the two members of the Metapenaeus genus. 12 random primers yielded a total of 82 fragments, of which, 50 bands were polymorphic whereas 32 fragments were monomorphic. Both molecular markers (RAPD and RFLP/PCR ) were facilitated the task of distinguishing species that are morphologically close species and very useful in determining the distribution of the exotic species.

Kappa-casein as a mammalian milk protein is involved in a several important physiological processes and it’s about 80% of the total protein in cow milk. This study aimed at genotyping bovine Kappa casein (CSN3) in two indigenous Nigerian cattle populations and to determine the frequency distribution of Kappa casein variants as detected across the animals examined and their association with the body measurement. DNA was extracted from 100 blood samples of 50 White Fulani and 50 N’dama cattle for identification and genotyping of kappa-casein gene by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) test using HindIII restriction endonucleases. The PCR products of the specific primer K-F and K-R for the two cattle breeds give 530bp specific band. Digestion of 530bp amplified products of White Fulani and N’dama by restriction endonuclease HindIII generated three fragments of 530-, 370- and 160- bp each for the two breeds. Results of the cuts with this enzyme show the presence of genotypes AA, AB and BB in the samples. These findings suggest that BB genotype could be selected for increase body conformation and protein content of milk. Olanrewaju, B. M. | Department of Animal Production and Health, The Federal University of Technology Akure (FUTA), Ondo State, Nigeria.

The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3 gene. The two most common gene variants in the HF breed are CSN3*A and CSN3*B while CSN3*E has been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A (0.584), CSN3*B (0.336) and CSN3*E (0.08). The observed genotype frequencies differed significantly (PF=0.17) indicated moderate heterozygosity deficiency. Nevertheless, the CSN3*B allele was present with relatively high frequency which should be used to positively select for its carriers, since increasing its frequency could help to improve the rheological properties of the milk intended for cheese production.

Cyanobacteria play a vital role in supplying nitrogen into the soil and aquatic ecosystem. It has an extra chromosomal DNA, whose role is not yet defined well. Isolation and characterization of extra chromosomal DNA in cyanobacteria might help to understand its survival mechanism. Cylindrospermum stagnale isolated (and deposited in NRMCF 3001) from soil showed presence of four plasmids namely pCYLM01, pCYLM02, pCYLM03, and pCYLM04. The following plasmids pCYLM01 and pCYLM02 were subjected to restriction digestion using HindIII restriction enzyme and cloned into pBlueScriptSK(-) vector. The sequence of pCYLM01 contained 4 potential open reading frames (ORFs) that have amino acids in the range of 59-299. Among them, ORF1 shows high sequence homology to the bacterial replication initiator family protein as evident from BLASTP analysis. The analysis of 4359 bp plasmid pCYLM02 sequence revealed 7 ORFs which are longer than 50 amino acids in length. The ORF2 of pCYLM02 has 243 amino acids and is represented in the plasmid sequence from 3045 to 3776 bp. The ORF3 of pCYLM02 corresponds to the plasmid sequence from 2323 to 2976 and codes for a putative protein of 217 amino acids long. A number of small ORFs below 50 bp were also found in the sequence analysis. Funding Agency: University of Algarve, Portugal info:eu-repo/semantics/publishedVersion

TOPARSLAN, Emine/0000-0001-8194-7842; MERCAN, Levent/0000-0002-6790-1458 WOS: 000500746600010 Thirty-five Anatolian water buffalo were selected from among a total of almost 7000 animals from 11 herds in the Kizilirmak Delta, which is known to be one of the most important areas for water buffalo farming in Turkey. The cleaved amplified polymorphic sequences (CAPS) method was used to identify single-nucleotide polymorphisms at 3 loci that are associated with milk production, namely PRL, CSN3, and PIT-1, which are associated with milk processing and milk yields, respectively. Sequence-tagged sites of PRL, CSN3, and PIT-1 were digested by restriction endonucleases RsaI, HindIII, and HinfI, respectively. Capillary electrophoretic analysis identified all samples as belonging to the GG and BB genotypes, which are undesirable genotypes for PRL and PIT-1 gene loci, but ideal for the CSN3 locus. Then the samples were sequenced by an ABI 3730 DNA Analyzer (Applied Biosystems). DNA sequencing of the amplified CSN3 fragment revealed one polymorphism at codon 135 (ThrACC > IleATC) in some samples. A homozygous structure was found for all three loci and the huge population loss between 1991 and 2008 suggests that a genetic bottleneck may have occurred in the population of Anatolian water buffalo in the Kizilirmak Delta.

Objective: To determine the association of Factor V Leiden / prothrombin gene mutation in Pakistani women with adverse pregnancy outcomes. Method: The prospective study was conducted at the Aga Khan University Hospital, Karachi, from January 1 to December 31, 2016, and comprised females ?40 years having history of two or more foetal losses with no apparent aetiology. Restriction fragment length polymorphism- Polymerase chain reaction was performed using MnlI and HindIII restriction enzymes for factor V Leiden G1691A and prothrombin gene mutation G20210A. Females with two or more consecutive normal pregnancies were enrolled as the control group. Data was analysed using SPSS 19. Results: Of the 172 participants with a mean age of 29.3±5.9 years (range: 19-38 years). 86(50%) each were healthy controls and those with recurrent pregnancy loss. There were 238 livebirths among the controls compared to 13 in the other group. Factor V Leiden G1691A was identified in 2(2.3%) women, and prothrombin gene mutation G20210A in 1(1.2%) woman in the patient group, while no mutation was identified in the control group. Conclusion: The prevalence of Factor V Leiden / prothrombin gene mutation in women with recurrent pregnancy loss was found to be very low. Continuous....

The published data indicate that the bacterium Clavibacter michiganensis subsp. sepedonicus (Cms) shows a notably low degree of intraspecific variation, and most of the molecular diagnostic methods could differentiate among subspecies groups but lacked the resolution to detect genetic diversity within subspecies. The present study describes effective differentiation of 50 isolates of C. michiganensis subsp. sepedonicus by the PCR MP method using five restriction enzymes (ApaI, PstI, BamHI, XmaI, HindIII). By analysing the electrophoretic patterns obtained by the PCR MP method performed with the applied restriction enzymes, each of the 50 tested isolates could be identified and distinguished from the remaining isolates. The PCR MP method of bacterial strain identification can be used in epidemiological studies, particularly for excluding the isolates of C. michiganensis subsp. sepedonicus as a source of primary infection.

Introduction. Multiple sclerosis is an inflammatory disease, where fibrin deposition and the impairment in its degradation have been shown to play an important role in the demyelination process. Tissue plasminogen activator (tPA) is a serine protease that enhances the conversion of plasminogen into its active form plasmin, the principal tPA inhibitor is the PAI-1. Several PAI-1 polymorphisms impact its gene expression and protein activity. Furthermore, the aim of this study was to investigate the association between the - 844 G>A, HindIII C>G, and 4G/5G PAI-1 polymorphisms and susceptibility to MS. Material and Methods. The study group included 400 Mexican mestizo subjects: 200 unrelated patients and 200 unrelated individuals identified as control subjects. The analysis of PAI-1 polymorphisms was performed by polymerase chain reaction-restriction fragment length polymorphism. Results. A significant association was found between the CG genotype of the HindIII C>G PAI-1 polymorphism and susceptibility to MS (OR=1.58, p=0.03); moreover, the frequency of 5G allele and 5G/5G genotype of the 4G/5G PAI-1 polymorphism was statistically significant (OR=1.36 and p=0.04 and OR=2.43 and p=0.02, respectively). With respect to the relation between the scores of progression (EDSS) and severity (MSSS), no association was found between EDSS and genotypes of the PAI-1 polymorphisms analyzed. Regarding MSSS, male that carries genotype GA of the -844 G>A and genotype 4G/5G of the 4G/5G PAI-1 polymorphisms showed a significant association with an increase of media of MSSS in comparison with females (p=0.01 in both cases).

Background Metabolic syndrome is a cluster of conditions that increase risk of cardiovascular morbidity and mortality. Among genetic factors that contributed to incidence of metabolic syndrome, Polymorphisms of Lipoprotein lipase (LPL) are major candidates especially because of their effect on obesity and dyslipidemia. S447X (rs328) and Hind III (rs320) Polymorphisms of LPL gene have been reported to change LPL activity, resulting in altered triglyceride (TG) and high density lipoprotein Cholesterol (HDL-C) levels. This study investigates the effects of these gene polymorphisms on factors affecting metabolic syndrome in northern population of Iran. Methods Studied population included 223 adults consisting 90 women and 133 men with body mass index (BMI) ≥ 30 kg/m2 as obese subjects, and 156 healthy participants as a control group with BMI Conclusions The present study proves that some genotypes of HindIII were associated with a reduced risk of developing low HDL-C only in men, while the protective effects of S447X on hypertriglyceridemia were only seen in women. The point is that this relation is affected by the weight profile of the participants. It can be concluded that there is a gender-related relation between the polymorphisms of LPL gene and the risk factors for incidence of metabolic syndrome in the northern population of Iran.

Groupers are biologically diverse, commercially important and highly regarded of all fish species associated with reef fisheries globally. Given the huge demand and high value in live fish trade, restaurants and supermarkets around the world, the accurate identification of the closely related grouper species is crucial to prevent any adulteration. Partial nucleotide sequence data of the mitochondrial cytochrome c oxidase I (COI) gene, which is globally accepted as fish DNA barcoding gene, was generated for six commercially important grouper species belonging to the genus, Epinephelus viz. E. fuscoguttatus, E. coioides, E. logispinis, E. malabaricus, E. bleekeri and E. tauvina collected from Indian waters. Based on the analysis of 704 bp COI gene sequences, a PCR-RFLP method was developed to identify all the six species accurately. The PCR products of COI gene fragment was digested with selected restriction enzymes viz. HinfI, AseI, HindIII and BsaJI. Restriction endonuclease HinfI was enabled to produce species-specific fragments for E. coioides, E. malabaricus, E. logispinis and E. bleekeri. The result shows that AseI restriction enzyme could be used to differentiate E. fuscoguttatus from other five species. The expected and actual molecular weight of the digested products was similar indicating the authenticity of the methods followed. This is the first report on differentiating closely related grouper species using above mentioned restriction endonucleases and COI gene fragment. PCR-RFLP profiles generated could be useful for detecting seafood fraud by identifying the grouper species under the genus Epinephelus using samples from fresh, frozen, cooked and even processed products.

In vivo control activities test of BCA candidate is required to evaluate its effectiveness. This test involved identification of the fungal species that inhabited the experimental plant after inoculation of BCA and pathogen. Identification of inhabitant fungi in the BCA control activity test can be conducted by culturing the fungi on artificial media. Fungal species can be identified based on morphological characters or genetic characters of fungal culture. This study was conducted to determine the potential of PCR ITS-RFLP molecular markers in the initial selection process of fungal isolates before identification based on DNA sequences and to select the enzyme that produce polymorphic PCR ITS RFLP pattern. Three enzymes (DraI, EcoRI and HinfI) were successfully separated 8 fungal cultures used in this study into three groups based on the pattern of PCR ITS DNA, while five other enzymes (BamHI, BclI HaeII, HpaI, and HindIII) were failed to cut the DNA ITS fragments, except for one isolate.

Background: Genetic information is necessary to devise strategic plans aimed to improve the genetic merit of buffalos. Objective: To assess the effect of genetic polymorphisms in GH, Pit-1, GHR, GHRHR, and KCN3 genes on milk production and body weight of Khuzestan water buffaloes. Methods: Blood samples were collected from 60 buffaloes from the Khuzestan province, Iran. Using the PCR-RFLP technique, the amplified and digested fragments of GH/AluI, GHR/AluI, GHRHR/ HaeIII, Pit1/HinfI, and KCN3/HindIII were genotyped. Results: All animals were monomorphic for GHRHR. The frequency of mutant alleles for GH, GHR, KCN3, and Pit1 was 47.5, 74.2, 49.2, and 51.7%, respectively. There were significant differences (p

Sutarno, Zahrah S, Astirin OP, Herawati E, Setyawan AD. 2019. Genetic diversity of Ongole Grade, Aceh, and Sumbawa cattle based on polymorphism on ND-5 fragment mitochondrial DNA using PCR-RFLP technique. Biodiversitas 20: 783-788. Genetic diversity is the basis of livestock breeding because it can be used as an initial improvement in livestock quality through artificial selection. This study aims to determine polymorphism in ND-5 fragment of mitochondrial DNA in Ongole Grade, Aceh, and Sumbawa cattle and their genetic diversity. The total DNA from the blood of the local cattle was extracted using the Wizard genomic DNA purification system from Promega and amplified using the PCR technique. The PCR product was then digested with HindIII enzyme using the RFLP technique to detect polymorphism. The genetic diversity of the Ongole Grade, Aceh, and Sumbawa cattle was analyzed using the formula from Nei and its genetic relationship was evaluated with the 2.02i NTSYSpc version program. Our findings showed there were polymorphisms in the ND-5 fragment of mitochondrial DNA. Digestion with HindIII restriction enzyme produces two types of haplotypes. Haplotype B is a 453 bp-sized DNA fragment that is not truncated by the HindIII enzyme, and haplotype A is a DNA fragment cut by HindIII enzyme into two with fragments of 336 bp and 117 bp. Polymorphism was found in Ongole Grade cattle, but not in Sumbawa and Aceh cattle. Haplotypes diversity in ND-5 fragments of mitochondrial DNA of Ongole Grade was 0.6250 while Sumbawa and Aceh cattle displayed no diversity of haplotypes. The genetic relationship shows that Sumbawa cattle belonged to the same cluster with Ongole Grade but separated from Aceh cattle.

Introduction: Beta-thalassemia is one of the most prevalent inherited blood diseases among Iranians. The aim of this study was to elucidate the chromosomal background of beta-thalassemia mutations in Esfahan province, Iran. Materials and Methods: In this study, we investigated three frequent mutations (c.315+1G>A, c.93-21G>A and c.92+5G>C in β-globin gene, the frequency of RFLP haplotypes, and LD between markers at β-globin gene cluster) in 150 beta-thalassemia patients and 50 healthy individuals. The molecular and population genetic investigations were performed on RFLP markers HindIII in the c.315+1G>A of Gγ (HindIIIG) and Aγ (HindIIIA) genes, AvaII in the c.315+1G>A of β-globin gene and BamHI 3' to the β-globin gene. All statistical analyses were performed using Power Marker software and SISA server. Results: Fifty percent of beta-thalasemia patients were associated with these mutations. Haplotype I was the most prevalent haplotype among beta-thalassemia patients (39.33%) and normal individuals (46%). The commonest c.315+1G>A mutation in our population was tightly linked with haplotype III (43.75%) and haplotype I (31.25%). The second prevalent mutation, c.92+5G>C, was 90%, 6.66%, and 3.33% in linkage disequilibrium with haplotypes I, VII, and III, respectively. The c.93-21G>A mutation indicated a strong association with haplotype I (80%). Conclusion: Our study participants like beta-thalassemia patients from Kermanshah province was found to possess a similar haplotype background for common mutations. The emergence of most prevalent mutations on chromosomes with different haplotypes can be explained by gene conversion and recombination. High linkage of a mutation with specific haplotype is consistent with the hypothesis that chromosomes carrying beta-thalassemia mutations experienced positive selection pressure, probably because of the protection against malaria experienced by beta-thalassemia carriers.

The comb is an accessory organ on the head of chicken which is influenced by testosterone hormone and can be used as an indicator of chicken’s fertility. Comb shape is related to climate adaptation and associated with a dominant mutation in chicken chromosome 1. Therefore this research was aimed to study the association between Pea-comb shape and SOX5 gene polymorphism in the population of progenies (BC-III Kambro) derived from a crossbreed between females Pelung and males second backcross generation Kampong Broiler-Type (BC-II Kambro). Chicken (Gallus gallus) SOX5 gene was acquired from NCBI GenBank with the Ref. Seq. 418195. Primers used to amplify the SOX5 gene are (F):5’-AGGTAGCCATGGTGACAAGC-3’,(R):5’-GATCTGTGAGGCAGCCAGTT-3’. Progenies showed 100% Pea-comb shape, while parental generation composed of Pea-comb shape and Single-comb shape. PCR-RFLP and endonuclease restriction enzyme HindIII were unable to determine the genotype of the female parent with Pea-comb. The result of SOX5 gene polymorphism showed the comb shape uniformity between progenies and parental of Kambro backcross generation. This study concluded that the genotype of Pea-comb shape female Pelung was undetermined and there was no polymorphism of the SOX5 gene between pea and single comb. PCR-RFLP using endonuclease restriction enzyme HindIII produced both target products and non-target/artifact products. The sequencing procedure was required to provide nucleotide sequences.

Background Sickle cell disease (SCD) is a common hematological genetic disorder in Saudi Arabia, Africa, the Mediterranean region, and India. The present study aimed to characterize βS haplotypes found in the Jazan region, Saudi Arabia. Methods One hundred sickle cell trait (SCT) individuals, diagnosed during their visit to the premarital screening clinic at King Fahad Central Hospital, were included in the study. Molecular analysis was carried out by polymerase chain reaction (PCR) and six polymorphic sites of the β-globin gene were analyzed using restriction endonucleases Hind II, Xmn-I, Hind III, and Ava II. Results The results of the current study revealed the presence of five typical haplotypes in which Benin, Bantu, and Senegal were found in homozygous state with 29%, 3% and 1% frequencies, respectively. Interestingly, 29% of the studied population showed atypical haplotypes in heterozygous state and 2% in homozygous state for the first time in Jazan region. Conclusions In addition to the typical haplotypes, high frequency of atypical haplotypes in this study indicates a diverse genetic mechanism that might have a crucial effect on the severity of SCD in this region. Therefore, considering this study in a cohort population with SCD in Jazan region may provide more indepth details about the correlation between haplotypes and the clinical manifestation of the disease.

Promoter-focused chromatin conformation techniques directly detect interactions between gene promoters and distal genomic sequences, providing structural information relevant to gene regulation without the excessive non-genic architectural data generated by full scale Hi-C. 3D promoter ‘interactome’ maps are crucial for understanding how epigenomic features like histone modifications and open chromatin, or genetic variants identified in genome wide association studies (GWAS), contribute to biological function. However, variation in sensitivity between such promoter-focused methods, principally due to restriction enzyme selection, has not been systematically assessed. Here, we performed a head-to-head comparison between promoter capture C (PCC) and promoter capture Hi-C (PCHiC) with the respective 4 cutters DpnII and MboI versus 6 cutter HindIII datasets from the same five cell types. While PCHiC generally produces a higher signal to noise ratio for significant interactions in comparison to PCC, we show that DpnII/MboI detects more proximal interactions and shows little overlap with the HindIII detection range. Promoter-interacting genomic regions mapped by 4-cutters are more enriched for regulatory features and disease-associated genetic variation than 6-cutters maps, suggesting that high-resolution maps better capture gene regulatory architectures than do lower resolution approaches.

Background: Lipoprotein lipase (LPL) plays an important role in plasma lipoprotein metabolism.An increasing number of studies have suggested an association of LPL gene variants with the risk ofcardiovascular and cerebrovascular diseases.Objective: To analyze whether Hind III polymorphism of LPL gene and serum LPL level are associatedwith ischemic stroke in Iraqi population.Method: Fifty ischemic stroke patients (clinical diagnosis and x-ray CT) and fifty controls were enrolled inthis case–control study. The LPL Hind III polymorphism was determined by PCR-RFLP technique and LPLmass level was estimated using a sensitive sandwich enzyme-linked immunosorbent assay (ELISA).Results: In the present research was not found any association between the Hindlll LPL gene polymorphismand acute ischemic stroke in the population studied; the allele and genotypic frequencies of the studiedpolymorphism was similar in cases and controls and followed the Hardy-Weinberg equilibrium. Therelationship between Hind III genotypes and the LPL mass level was analyzed using ANOVA and furtherconfirmed by Post-hoc analysis. there was no significant difference in LPL mass levels between the genotypegroups H+H+, H+H-, and H-H- (p value>0.05).Conclusions: The Hind III polymorphism of LPLis not a genetic marker for the development of ischemicstroke as well as not determinants of serum LPL level in the Iraqian sample used.

BACKGROUND Genetic predisposition is an important risk factor in coronary artery disease (CAD).This study was conducted to determine the polymorphism frequencies of the plasminogen activator inhibitor-1(PAI-1) gene 4G/5G, Angiotensin-converting enzyme (ACE) gene I/D,and Angiotensin II type 1 receptor (AT1) gene A1166C genotypes and to examine the role of these polymorphisms in CAD. METHODS Genomic DNAs obtained from 260 subjects(130 CAD patients and 130 control) were used in the study. ACE I/D and PAI-1 4G/5G polymorphism genotypes were determined using polymerase chain reaction (PCR) and electrophoresis. AT-1 A1166C polymorphism was determined using the PCR, restriction fragment length polymorphism (RFLP) and electrophoresis. The products amplified from AT1 gene by PCR were cut with HindIII restriction endonuclease and then analyzed by 2% agarose gel electrophoresis. The results were statistically analyzed with the chi-square test, Mann-Whitney U test, and independent two-sample t-test. RESULTS Allele frequencies showed statistically significant differences between the patient and control groups. There was no statistically significant difference in ACEI/D genotype frequencies between the twogroups. Likewise, no statistically significant difference was found in the AT1 A1166C genotype frequencies; however, a statistically significant difference was found in allele frequencies.The PAI-1 4G/5G genotype frequency was significantly higher in the patient group. CONCLUSIONS While there is a relationship between of PAI-1 gene 4G/5G polymorphism and CAD, ACE gene I/D and AT1 gene A1166C polymorphisms are not related. PAI-1 gene homozygous genotypes may be considered as a prognostic marker for CAD patients.

Porcine circovirus type 3 (PCV3), which currently lacks effective preventive measures, has caused tremendous economic losses to the pig husbandry. Obtaining the strain of PCV3 is the key to preparing related vaccines and developing corresponding antiviral drugs. In this study, according to the linear sequence of PCV3 DNA published on GenBank, the sequence was rearranged with SnapGene gene-editing software, and after rearrangement, the HindIII restriction endonuclease site was added to the end of the linear DNA, so that both ends have the same restriction endonuclease site. On this basis, the rearranged linear DNA is obtained by gene synthesis, PCR amplification, DNA purification, etc., and is digested and connected in vitro to obtain cyclized DNA. PCV3 infectious clones were obtained by transfecting 3D4/21 cell lines. The obtained PCV3 was identified by PCR, Western blotting, and indirect immunofluorescence tests. The results showed that this study successfully obtained the strain of PCV3 in vitro. To further evaluate the pathogenicity of the obtained PCV3 infectious clones, this study established an animal model of Kunming mice infected with PCV3. The results of RT-PCR, Western blotting and immunohistochemistry showed that PCV3 can infect myocardium and alveoli of Kunming mice, but no PCV3 was detected in other tissues. The above studies indicate that PCV3 circular DNA can be used to construct PCV3 infectious clones. This research will provide a new method for the construction of circular DNA viruses and lay the foundation for the research and pathogenesis of PCV3 vaccine.

To evaluate the role of erythrocyte complement receptor 1 (ECR1) gene in the predisposition to respiratory distress syndrome (RDS), we studied 50 infants with RDS and 50 controls. Real-time polymerase chain reaction allelic discrimination analysis of A3650G (rs2274567) and genotyping of the alleles (HindIII) were performed. Allele L of HindIII restricted single nucleotide polymorphism (SNP) associated with the severity of RDS. Duration of oxygen and ventilation in genotypes AA and AG of A3650G SNP was longer than genotype GG (17.6 ± 19.4 and 8.6 ± 4.5 days, p = 0.01) and (8.9 ± 11.9 and 3.9 ± 3.53 days, p = 0.03), respectively. A3650G and HINDIII digested gene polymorphisms of ECR1 may be of little importance for RDS.

Bovine herpesvirus 1 (BoHV-1) causes several clinical syndromes in cattle worldwide. There are 3 subtypes of BoHV-1: 1.1, 1.2a, and 1.2b. Several molecular methods are commonly used in the detection and characterization of BoHV-1. Among them, restriction endonuclease analysis (REA) and single-nucleotide polymorphism (SNP) analysis of the complete viral genome allow classification of BoHV-1 into different subtypes. However, developing countries need simpler and cheaper screening assays for routine testing. We designed a standard multiplex PCR followed by a REA assay allowing straightforward subclassification of all BoHV-1 isolates tested into 1.1, 1.2a, and 1.2b subtypes based on the analysis of fragment length polymorphism. Our standard multiplex PCR-REA was used to analyze 33 field strains of BoHV-1 isolated from various tissues. The assay confirmed the subtype identified previously by REA. In addition, non-polymorphic or undigested fragments were sequenced in order to confirm the mutation affecting the RE HindIII site. Our PCR-REA method is an affordable and rapid test that will subtype all BoHV-1 strains. Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Virología E Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Virología E Innovaciones Tecnológicas; Argentina Fil: Miño, Orlando Samuel. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Apóstolo, Romina María. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: De Stefano, Gabriel A.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Romera, Sonia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Virología E Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Virología E Innovaciones Tecnológicas; Argentina

Background. Gardnerella vaginalis, a microorganism highly linked to bacterial vaginosis (BV), is understudied in terms of genotypic heterogeneity in South African populations. This study investigated the prevalence of G. vaginalis genotypes in BV-positive, BV-intermediate, and BV-negative South African pregnant women. Methods. The study population included n=354 pregnant women recruited from a public hospital in Durban, South Africa. The women provided self-collected vaginal swabs for BV diagnosis by Nugent scoring. For the genotyping assays, the 16S rRNA and sialidase A genes from BV-negative, BV-intermediate, and BV-positive samples were amplified with G. vaginalis-specific primers. The16S rRNA amplicon was digested with TaqI to generate genotyping profiles, and subtypes were determined by correlating BamHI and HindIII digestion profiles. Phylogenetic analysis was performed on the 16S rRNA and sialidase A sequences. The data analysis was performed with R Statistical Computing software, version 3.6.2. Results. Two different genotypes, GT1 and GT2, were detected. The most prevalent genotype was GT1. Four subtypes (1, 2B, 2AB, and 2C) were shown to be present. The most prevalent subtype was 2B, followed by subtypes 1, 2C, and 2AB. The phylogenetic analysis of the 16S rRNA showed the presence of 5 clusters. The tree displayed clusters which contained sequences from the same BV group with different genotypes and subtypes. Clusters with sequences from across the BV groups carrying the same genotype and subtype were present. Diversity of the sialidase A across BV groups and genotypes was observed. Finally, the study did not find a significant association (p>0.05) between reported symptoms of abnormal vaginal discharge and genotype harboured. Conclusion. This study provided the first report on the diversity of G. vaginalis in South African pregnant women. Diversity assessments of G. vaginalis with respect to genotypes and virulence factors may aid in a greater understanding of the pathogenesis of this microorganism.

B-cell non-Hodgkin lymphoma (B-NHL) are clonal malignancies with diverse clinical presentations, pathogenesis, and biologic behavior. Hepatitis C virus (HCV) is a major leading cause of liver-related morbidity and B-cell lymphoproliferative diseases such as mixed cryoglobulinemic syndrome (MCns) and B-NHL. Fibronectin (FN) is a multifunctional glycoprotein of 250 kDa which is encoded on a gene that is 75 kb in length, located on chromosome 2q34–36 and consists of 50 exons. Polymorphisms of the fibronectin gene are as follows: HindIII, HaeIII, TaqI, and MspI. The aim of the current study was to investigate the possible link between the two FN polymorphisms (called MspI “CC genotype, CD genotype and DD genotype” and HaeIII “AA genotype, AB genotype and BB genotype”) and the development of lymphoma in HCV-positive patients with mixed cryoglobulinemic syndrome compared to healthy age- and sex-matched individuals. The present study included 75 patients; 25 HCV-positive patients with MCns and 50 B-NHL patients, in addition to 25 healthy controls. To achieve this aim, genotyping for FN gene was done by PCR-RFLP technique and screening for HCV infection was performed by ELISA and confirmed by RT-PCR, cryoglobulins have been detected by cold precipitation (4 °C for 72–96 h) and plasma FN levels have been assessed by ELISA. The study revealed that HaeIII-AB genotype can be considered as a risk factor for NHL development and high HaeIII-A allele frequency seems to be associated with NHL development. We did not find any significant difference between all patients’ groups and controls regarding MspI genotyping. It was also found that plasma FN level did not show any significant difference between B-NHL patients and each of HCV positive patients with MCns and controls. Our study provides further evidence for the role of HaeIII-AB FN gene polymorphism as a risk factor for B-NHL development in sub-population of Egyptian HCV-positive patients with MCns. We also found that HaeIII-A allele has been significantly increased in overall B-NHL patients.

Red stripe caused by the bacterium Acidovorax avenae subsp. avenae (Aaa) is a disease of sugarcane that is distributed worldwide. In this study, 108 sugarcane leaf samples were collected in 2013–2016 from nine sugarcane‐growing regions in China. Aaa was detected by PCR with specific and novel primers from the 16S–23S rDNA internal transcribed spacer region in 81 of 84 (96%) leaves with red stripe symptoms and in 20 of 24 (83%) leaves without symptoms. Furthermore, Aaa was detected in all nine sampling locations representing six sugarcane‐producing provinces in China. The 101 amplified fragments were cloned and sequenced. The size of the nucleotide sequences varied from 436 to 454 bp and the sequence identity ranged from 89.2% to 100%, suggesting a significant genetic variation among Aaa strains from China. Five major restriction fragment length polymorphism (RFLP) profiles were obtained by in silico and polyacrylamide gel electrophoresis analyses of the PCR products digested with HindIII and EcoRI. The causal agent of sugarcane red stripe was also successfully isolated from a diseased plant and its pathogenicity confirmed by inoculation of healthy sugarcane plantlets and reproduction of disease symptoms. The data showed that Aaa is currently widespread in China, suggesting that control methods should be implemented to limit the impact of red stripe on sugarcane production.

Due to the feature of high hydrolysis, tannase is widely used in food, beverage, brewing and other fields. However, high cost in producing natural tannase makes it difficult to apply tannase to industry in a large-scale. Microbial expression systems can be used for preparing numerous amount of enzyme at low cost, so in this paper Aspergillus niger N5-5 was expressed using E. coli system. Specific primers were designed based on the Aspergillus niger N5-5 sequence N3 (GenBank, No.: KP677552), and tannase gene tan was promoted to carry 6 His tag and enzyme cutting site which contains NdeI/HindIII using PCR amplification. Then, tannase gene tan was connected to expression vector by NdeI/HindIII enzyme cutting. In this way, recombinant expression vector tan-pET43.1a was formed. Then, the expression vector pET43.1a by NdeI/HindIII enzyme cutting was transformed into E. coli BL21 (DE3) to induce expression of Aspergillus niger N5-5. When the induced fungi were disrupted by the ultrasonic wave, the crude enzyme was extracted and purified by using the IMAC, and then the activity of the crude enzyme and pure enzyme was determined. According to the results of determination of the tannase activity, the tannase activity of the crude enzyme was greatly improved after the crude enzyme was purified, and the specific activity of the pure enzyme was about 8 times of that of the crude enzyme. The results of SDS-PAGE of the pure enzyme showed that the molecular mass of the pure enzyme was about 65 kDa/64–65 kDa, which was consistent with the expected result (64.2 kDa), It can be concluded that the crude enzyme solution was purified successfully. The results of pure enzyme’s protein identification by Western Blotting showed that clear protein bands pro-3 were observed. Molecular mass of clear protein bands pro-3 was about 65 kDa, which was in line with the expected results (64.2 kDa). It can be seen that the aforementioned expression protein could be specifically combined with His tag. It proved expression protein to be a recombinant fusion protein with 6 His tag.

Heterochromatin in the genome plays a vital role in species differentiation and evolution. To characterize and unravel molecular composition of the heterochromatic regions of the genome in dipteran flies, especially in Sarcophagids, different banding techniques have been used previously which established the banding pattern of constitutive heterochromatin. The present study is an attempt to further explore the base specificity of rather highly condensed C-banded regions of chromosomes using restriction enzyme (RE) digestion along with Giemsa staining, in search of species-specific targets of RE digestion. Different REs such as Apa I, Cla I, Eco RI, Hind III, Hae III and Pvu II have been used to characterize the heterochromatic regions of two Sarcophaga species, i.e., Sarcophaga ruficornis and Sarcophaga argyrostoma. Some of the constitutively heterochromatic regions remain intensely stained indicating no targets for the enzyme cleavage, while some of the heterochromatic areas in both the species show remarkable and distinct digestion sites after the treatment of REs. In S. ruficornis, Hind III digests the entire autosomes including the pericentromeric regions and the sex chromosome; while Hae III and Pvu II selectively digest autosomes and sex chromosome. While in S. argyrostoma, Cla I digests the entire autosomes including the pericentromeric regions; Hind III and Hae III selectively digests autosomes and sex chromosomes. These findings suggest the presence of distinctive enzyme digestion targets on the heterochromatin, which may provide important evidence for the differential base specificity that were previously found to be constitutive in nature.

BACKGROUND Several studies have studied the relationship between lipoprotein lipase (LPL) HindIII gene polymorphism and stroke susceptibility. However, the conclusions remain controversial. To clarify the association of LPL gene HindIII polymorphism and stroke susceptibility, we therefore conducted a comprehensive meta-analysis. MATERIALS AND METHODS The PubMed, Web of Science, EMBASE, and Google Scholar databases were systemically searched to indentify available studies. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated under the allelic, dominant, homozygous, heterozygous, and recessive models. The data were analyzed by using Stata 12.0 (StataCorp, College Station, TX). RESULTS Ten studies were enrolled, including a total of 2122 cases and 2235 controls. The overall results showed that LPL HindIII variants were associated with a decreased risk of stroke (G versus T: OR = .78, 95% CI = .70-.87, P

We conducted a cross-sectional study to investigate the effects of the adenosine triphosphate-binding cassette transporter 1 (ABCA1) I883M and lipoprotein lipase (LPL) HindIII polymorphisms on lipid levels in patients with hyperlipidemia. A total of 533 patients were enrolled. Serum lipid parameters were determined by an automatic biochemistry analyzer. Genotyping of the ABCA1 I883M and LPL HindIII was carried out using the polymerase chain reaction-restriction fragment length polymorphism technique. Multiple linear regression analysis was used to estimate the associations between serum lipid levels and the genetic polymorphisms. The frequency distribution of the ABCA1 I883M and LPL HindIII polymorphisms did not deviate from Hardy-Weinberg equilibrium. The major finding of our regression analysis showed that neither the ABCA1 I883M nor the LPL HindIII polymorphism was associated with baseline serum lipid levels in the total population. However, among patients with elevated alanine aminotransferase (ALT) levels (ALT ≥ 40 U/L), carriers of the M allele of the ABCA1 gene had lower levels of high-density lipoprotein cholesterol (HDL-C) and higher levels of low-density lipoprotein cholesterol (LDL-C) after adjusting for age, sex, smoking status, alcohol consumption, education level, occupation, and work intensity ( P < .05 for both). A test on interaction terms between the ABCA1 I833M polymorphism and ALT on HDL-C and LDL-C levels also remained significant ( P = .001 and P = .014, respectively). Our data suggest that there are significant interactive effects between ABCA1 I883M and ALT levels on HDL-C and LDL-C levels. However, the LPL HindIII polymorphism did not influence lipid levels.

Gurami ( Osphronemus gouramy Lac.) is a popular fish species among Indonesian people. Several Gurami strains have been cultivated by fish farmer, one of which is Gurami Soang. This strain is belived to have a faster growth rate compared to other strains. However, like other strains, the fingerling of Soang strain have also a low survival and suceptible to disease, especially that caused by A eromonas hydrophila infection. It has been proved that seeds from a single spawning event show varibale disease resistance. The difference in resistance among individuals is suggested related to the difference in their genetic component. One of the genes responsible for resistance is Major Histocompatibility Complex ( MHC ) class II B gene. Variability in resistance can be analyzed by using PCR - RFLP technique. PCR-RFLP is a technique that can produce a specific DNA fragments by PCR, followed by cutting the PCR product using restriction enzymes to describe the presence or absence of restriction sites in DNA fragments. This research aims to determine genetic marker to differiantiate between resitant and irresistant individual of Gurami Soang infected by A. hydrophila which die at a different time priod based on PCR-RFLP MHC class IIB gene. The study used survey method with purposive random sampling. The Data of PCR-RFLP band patterns were analyzed descriptively. The result indicated that cutting of the MHC class II B gene using Hinf I produce two RFLP bands with 300 bp and 100 bp length in all samples. Meanwhile, the MHC IIB gene was not cuted by Pst I, Hind III, Bam HI and Eco RI enzymes forall samples. These mean that MHC II gene in all individuals were monomorphic. Therefore,it can be concluded that there is no specific genetic marker to differentiate gurami soang individulas which was dying in different time periods.

The gypsy moth (Lymantria dispar L., Lepidoptera: Lymantriidae) is one of the most serious pest of various forestal, food and industrial crops worldwide. We have characterized a new Lymantria dispar nucleopolyhedrovirus (LdMNPV-T4) variant, which was isolated from dead L. dispar larvae in Turkey. Scanning electron microscope observations showed that the polyhedral occlusion bodies (OBs) of the LdMNPV-T4 were irregularly shaped. Transmission electron microscopy revealed that OBs of LdMNPV-T4 were occupied with several virions in which multiple nucleocapsids packaged by viral envelope. Restriction analysis of the LdMNPV-T4 DNA purified from the viral inclusion bodies yielded BamHI, BglII, EcoRI and HindIII fragments. The mean size estimated for the complete LdMNPV-T4 genome was calculated to be 163.3 kb. Phylogenetic analysis of amplified polh, lef-8 and lef-9 sequences showed its relation to the other NPVs from Lymantria species. Mortality values of the LdMNPV-T4 at four different concentrations ag...

Background Lipoprotein lipase (LPL) is one of the major enzymes responsible for the hydrolysis of triglyceride (TG)-rich lipoprotein. The effects of LPL polymorphisms on serum TG are inconsistent among different populations. Objectives This study aims to assess the TG serum concentration and distributions of three LPL single nucleotide polymorphisms (SNPs), namely D9N, HINDIII and S447X, in a nationally representative sample of Iranian adolescents. Material and methods We studied the associations between SNP genotypes and TG levels in a nationally representative sample of Iranian adolescents. Genotyping was performed in 750 randomly selected participants. We compared the genotypes according to different TG levels. Results This study comprised 746 participants, with mean ± SD age of 14.6 ± 2.5 years. The distribution of genotypes of D9N and S447X were not significantly different according to TG levels. Regarding the HINDIII polymorphism, the distribution of GG, GT, and TT genotypes were significantly different in participants with low, borderline-high, and elevated TG (p = 0.02, 0.03, and 0.01, respectively). The mean TG was not significantly different according to the genotype distribution. Conclusions In this study, most of the LPL gene variants were not significantly different in adolescents with normal and elevated TG, and the mean TG was not different in participants with various genotypes. As the first evidence from the pediatric population of the region of the Middle East and North Africa (MENA), these results might be used in international comparisons. Our findings might suggest that the high prevalence of hypertriglyceridemia in Iranian adolescents is more likely to be a result of lifestyle rather than genetic factors.

SUMMARYPolymorphisms in the myostatin (MSTN) gene were detected in 150 female Iranian Markhoz goats. Two 573 base pairs (bp) and 475 bp fragments of the MSTN gene, which contains a deletion 5 bp indel (206 TTTTA/), in the region of exon 1 encoding the 5′ untranslated region (UTR) of the MSTN transcript, and two single-nucleotide polymorphisms (SNPs) of substitution (339T/A, 169T/G) in exon 1 and 3 regions, respectively, were amplified. The polymerase chain reaction (PCR) products were digested separately using restriction enzyme endonuclease DraI, HinIII and HindIII. The digestion results indicated AA and AB genotypes in the region of exon 1 encoding the 5′UTR of the MSTN transcript, AA, AT and TT genotypes in exon 1 and TT, TG and GG genotypes in exon 3. The SNPs loci were in Hardy–Weinberg disequilibrium but the deletion locus showed equilibrium in the Markhoz goat population. Evaluation of associations between the polymorphisms with the studied growth traits showed that the AA and GG genotypes of exons 1 and 3 have a significant positive effect on weight at 6 months of age (W6) and average daily gain (ADG) traits, but genotypes in the region of exon 1 encoding the 5′UTR of the MSTN transcript did not have any significant effect on the studied growth traits. The statistical analyses showed a positive and significant effect of the 339A allele (exon 1) for W6 and negative and significant effect of the 169G allele (exon 3) for ADG trait. Therefore, these results suggest that the MSTN gene could be a potential candidate gene that affects ADG and W6 traits in goats. More studies are needed to simultaneously consider variants of this region in a larger population to better understand MSTN gene effects on the economic traits in goat.

Type 2 diabetes mellitus (T2DM) belongs to the diseases with hereditary predisposition, so both environmental and genetic factors contribute to its development. Recent studies have demonstrated that the skeleton realizes systemic regulation of energy metabolism through the secretion of osteocalcin (OCN). Thus, the association analysis between HindIII single nucleotide polymorphism of OCN gene (BGLAP) promoter region and T2DM development in Ukrainian population was carried out. 153 individuals diagnosed with T2DM and 311 control individuals were enrolled in the study. The genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The lack of association between BGLAP HindIII single nucleotide polymorphism (SNP) and T2DM development among Ukrainians was found. Further studies with extended groups of comparison are needed to confirm the obtained results.