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Background Segmented filamentous bacteria (SFB) are intestinal commensal microorganisms that have been demonstrated to induce the innate and adaptive immune responses in mouse and rat hosts. SFB are Gram-positive, spore-forming bacteria that fail to grow optimally under in vitro conditions due to unique metabolic requirements. Recently, SFB have been implicated in improved health and growth outcomes in commercial turkey flocks. To assess the nature and variations in SFB of turkeys and how they may differ from mammalian-associated SFB, the genome of turkey-associated SFB was compared with six representative genomes from murine hosts using an in silico approach. Results The SFB-turkey genome is 1.6 Mb with a G + C content of 26.14% and contains 1,604 coding sequences (CDS). Comparative genome analyses revealed that all the seven SFB strain possesses a common set of metabolic deficiencies and auxotrophies. Specifically, the inability of all the SFB strains to synthesize most of the amino acids, nucleotides and cofactors, emphasizing the importance of metabolite acquisition from the host intestinal environment. Among the seven SFB genomes, the SFB-turkey genome is the largest and contains the highest number of 1,604 predicted CDS. The SFB-turkey genome possesses cellular metabolism genes that are absent in the rodent SFB strains, including catabolic pathways for sucrose, stachyose, raffinose and other complex glycans. Other unique genes associated with SFB-turkey genome is loci for the biosynthesis of biotin, and degradation enzymes to recycle primary bile acids, both of which may play an important role to help turkey associated SFB survive and secure mutualism with its avian host. Conclusions Comparative genomic analysis of seven SFB genomes revealed that each strain have a core set of metabolic capabilities and deficiencies that make these bacteria challenging to culture under ex vivo conditions. When compared to the murine-associated strains, turkey-associated SFB serves as a phylogenetic outgroup and a unique member among all the sequenced strains of SFB. This turkey-associated SFB strain is the first reported non-mammalian SFB genome, and highlights the impact of host specificity and the evolution of metabolic capabilities.

Graphical abstract COVID-19, the disease caused by SARS-CoV-2, has been declared as a global pandemic. Traditional medicinal plants have long history to treat viral infections. Our in silico approach suggested that unique phytocompounds such as emodin, thymol and carvacrol, and artemisinin could physically bind SARS-CoV-2 spike glycoproteins (6VXX and 6VYB), SARS-CoV-2 B.1.351 South Africa variant of Spike glycoprotein (7NXA), and even with ACE2 and prevent the SARS-CoV-2 binding to the host ACE2, TMPRSS2 and neutrapilin-1 receptors. Since Chloroquine has been looked as potential therapy against COVID-19, we also compared the binding of chloroquine and artemisinin for its interaction with spike proteins (6VXX, 6VYB) and its variant 7NXA, respectively. Molecular docking study of phytocompounds and SARS-CoV-2 spike protein was performed by using AutoDock/Vina software. Molecular dynamics (MD) simulation was performed for 50ns. Among all the phytocompounds, molecular docking studies revealed lowest binding energy of artemisinin with 6VXX and 6VYB, with Etotal −10.5 KJ mol−1 and −10.3 KJ mol−1 respectively. Emodin showed the best binding affinity with 6VYB with Etotal −8.8 KJ mol−1and SARS-CoV-2 B.1.351 variant (7NXA) with binding energy of −6.4KJ mol−1. Emodin showed best interactions with TMPRSS 2 and ACE2 with Etotal of −7.1 and −7.3 KJ mol−1 respectively, whereas artemisinin interacts with TMPRSS 2 and ACE2 with Etotal of −6.9 and −7.4 KJ mol−1 respectively. All the phytocompounds were non-toxic and non-carcinogenic. MD simulation showed that artemisinin has more stable interaction with 6VYB as compared to 6VXX, and hence proposed as potential phytochemical to prevent SARS-CoV-2 interaction with ACE-2 receptor. Supplementary Information The online version contains supplementary material available at 10.1007/s40495-021-00259-4.

BackgroundSoil salinity has been one of the biggest hurdles in achieving better crop yield and quality. Plant growth-promoting rhizobacteria (PGPR) are the symbiotic heterogeneous bacteria that play an important role in the recycling of plant nutrients through phytostimulation and phytoremediation. In this study, bacterial isolates were isolated from salt-polluted soil of Jhajjar and Panipat districts of Haryana, India. The potential salt-tolerant bacteria were screened for their PGPR activities such as phosphate solubilization, hydrogen cyanide (HCN), indole acetic acid (IAA) and ammonia production. The molecular characterization of potent isolates with salt tolerance and PGPR activity was done by 16S rDNA sequencing.ResultsEighteen soil samples from saline soils of Haryana state were screened for salt-tolerant bacteria. The bacterial isolates were analyzed for salt tolerance ranging from 2 to 10%. Thirteen isolates were found salt tolerant at varied salt concentrations. Isolates HB6P2 and HB6J2 showed maximum tolerance to salts at 10% followed by HB4A1, HB4N3 and HB8P1. All the salt-tolerant bacterial isolates showed HCN production with maximum production by HB6J2. Phosphate solubilization was demonstrated by three isolates viz., HB4N3, HB6P2 and HB6J2. IAA production was maximum in HB4A1 (15.89) and HB6P2 (14.01) and least in HB4N3 (8.91). Ammonia production was maximum in HB6P2 (12.3) and least in HB8P1 (6.2). Three isolates HB6J2, HB8P1 and HB4N3 with significant salt tolerance, and PGPR ability were identified through sequencing of amplified 16SrRNA gene and were found to beBacillus paramycoides,Bacillus amyloliquefaciensandBacillus pumilus, respectively.ConclusionsThe salt-tolerant plant growth-promoting rhizobacteria (PGPR) isolated from saline soil can be used to overcome the detrimental effects of salt stress on plants, with beneficial effects of physiological functions of plants such as growth and yield, and overcome disease resistance. Therefore, application of microbial inoculants to alleviate stresses and enhance yield in plants could be a low cost and environmental friendly option for the management of saline soil for better crop productivity.

The aim of the present study was to test the binding affinity of methylxanthines (caffeine/theine, methylxanthine, theobromine, theophylline and xanthine) to three potential target proteins namely Spike protein (6LZG), main protease (6LU7) and nucleocapsid protein N-terminal RNA binding domain (6M3M) of SARS-CoV-2. Proteins and ligand were generated using AutoDock 1.5.6 software. Binding affinity of methylxanthines with SARS-CoV-2 target proteins was determined using Autodock Vina. MD simulation of the best interacting complexes was performed using GROMACS 2018.3 (in duplicate) and Desmond program version 2.0 (academic version) (in triplicate) to study the stabile interaction of protein-ligand complexes. Among the selected methylxanthines, theophylline showed the best binding affinity with all the three targets of SARS-CoV-2 (6LZG - 5.7 kcal molThe online version contains supplementary material available at 10.1007/s40495-021-00276-3.

Low temperature (LT) causes significant yield losses in chickpea (Cicer arietinum L.). The sucrose starch metabolism is associated with abiotic-stress tolerance or sensitivity in plants. The changes in sugars and starch contents under LT in chickpea have already been studied, however, no information is available on LT-induced alterations in transcription of carbohydrate metabolic pathway genes in chickpea. To understand the differences in the regulation of sucrose and starch metabolism under LT, the expression of sucrose and starch metabolism genes was studied in leaves of cold-sensitive (GPF2) and cold-tolerant (ICC 16349) chickpea genotypes. The mRNA sequences of chickpea genes were retrieved from the public databases followed by confirmation of identity and characterization. All the genes were functional in chickpea. Between the two paralogues of cell wall invertase, cell wall invertase 3×2 (CWINx2) was the truncated version of cell wall invertase 3×1 (CWINx1) with the loss of 241 bases in the mRNA and 67 amino acids at N terminal of the protein. Comparison of expression of the genes between control (22°C day / 16°C night) and LT treated (4°C; 72 h) plants revealed that granule bound starch synthase 2 (GBSS2) and β-amylase 3 (BAM3) were upregulated in ICC 16349 whereas sucrose phosphate synthase 2 (SPS2), CWINx1, CWINx2 and β-amylase 1 (BAM1) were downregulated. In contrast to this, SPS2, CWINx1, CWINx2 and BAM1 were upregulated and GBSS2 downregulated in GPF2 under LT. The gene expression data suggested that UGPase, CWINs, GBSS2 and BAM3 are important components of cold-tolerance machinery of chickpea.

Background Eukaryotic initiation factor 2B (eIF2B) initiates and regulates translation initiation in eukaryotes. eIF2B gene mutations cause leukoencephalopathy called vanishing white matter disease (VWM) in humans and slow growth (Slg−) and general control derepression (Gcd−) phenotypes in Saccharomyces cerevisiae. Results To suppress eIF2B mutations, S. cerevisiae genomic DNA library was constructed in high-copy vector (YEp24) and transformed into eIF2B mutant S. cerevisiae strains. The library was screened for wild-type genes rescuing S. cerevisiae (Slg−) and (Gcd−) phenotypes. A genomic clone, Suppressor-I (Sup-I), rescued S. cerevisiae Slg− and Gcd− phenotypes (gcd7-201 gcn2∆). The YEp24/Sup-I construct contained truncated TAN1, full length EMC4, full length YGL230C, and truncated SAP4 genes. Full length EMC4 (chaperone protein) gene was sub-cloned into pEG (KG) yeast expression vector and overexpressed in gcd7-201 gcn2∆ strain which suppressed the Slg− and Gcd− phenotype. A GST-Emc4 fusion protein of 47 kDa was detected by western blotting using α-GST antibodies. Suppression was specific to gcd7-201 gcn2∆ mutation in eIF2Bβ and Gcd1-502 gcn2∆ in eIF2Bγ subunit. Emc4p overexpression also protected the wild type and mutant (gcd7-201 gcn2∆, GCD7 gcn2∆, and GCD7 GCN2∆) strains from H2O2, ethanol, and caffeine stress. Conclusions Our results suggest that Emc4p is involved in eIF2B-mediated translational regulation under stress and could provide an amenable tool to understand the eIF2B-mediated defects.

Chickpea has a narrow genetic base, and consequently many DNA-based markers were unsuccessful in revealing polymorphism. The problem was overcome by the development of simple sequence repeat and single nucleotide polymorphism markers that revealed adequate polymorphism and these became markers of choice in chickpea. In addition, transcription factor gene–derived microsatellite markers, intron targeted primers, expressed sequence tag polymorphisms, diversity arrays technology, and insertion-deletion markers have also been developed for chickpea. Genotyping by sequencing is also gaining popularity. Saturated linkage maps of chickpea with adequate marker density on each linkage group have also been constructed; however, several of the recently developed markers are still to be placed on linkage maps. The markers have subsequently been used to map genes or identify quantitative trait loci (QTLs) for biotic stress resistance, abiotic stress tolerance, and agro-morphological traits. Using reference genomes of chickpea and QTL intervals, candidate genes in QTL regions for several of these characters have been identified. Notably, genes involved in flowering in chickpea have been identified. Same QTL was also found to govern more than one trait, e.g., drought tolerance and plant vigor, salinity tolerance and higher plant vigor, salinity tolerance and higher reproductive success. The markers linked closely to genes/QTLs of interest have also been exploited to introgress wilt and blight resistance, and double podding character into agronomically superior cultivars.

“Candidatus Arthromitus” UMNCA01 was recovered from ileal samples of commercial turkey poults and may have probiotic capabilities. The complete genome was determined using the Illumina MiSeq and HiSeq sequencing platforms. The complete genome consists of 1,631,326 bp and has a G+C content of 26.14%, 1,540 coding sequences (CDS), and 37 RNA coding genes.

Terminalia arjuna Wight & Arn is used in the Indian traditional medicinal system for the treatment of various cardiac disorders and has been used for the treatment of various human problems like blood diseases, anemia, viral disease, fractures, ulcers, hepatic problems. This study was designed to understand the genetic variation in T. arjuna from different geographical regions of North-west Himalayas. Using the RAPD-PCR method, a total of 183 DNA bands were amplified from 30 accessions of T. arjuna with an average of 6.1 bands per primer. The amplified band size ranges from 150-1700 bp. The Polymorphic Information Content (PIC) value ranges from 0.244 (OPB-10) to 0.781 (OPA-02). The Marker Index (MI) values ranged from 0.6075 to 6.248, with an average of 3.1378. The effective multiplex ratio (EMR) values range from 1.33 (OPC-12) to 9 (OPC-20). A positive correlation was observed between PIC and MI (r = 0.83), MI and EMR (r = 0.873) values. The percentage of polymorphic loci among T. arjuna populations was found to be 71.17%, while AMOVA showed 85% variance within the populations and 15% among the populations. From UPGMA cluster analysis, intermixing of accessions of different populations was observed. This study concludes a genetic variation among different accessions of T. arjuna, indicating good cross-pollination, leading to gene flow between closely located accessions without affecting the gene pool. The genetic diversity information of T. arjuna reported in this study is a very important component for efficient conservation and effective management of its genetic resources.

We report here the genome sequence of halophilic Halobacillus trueperi SS1, isolated from the Lunsu saltwater body in India. The bacteria are Gram positive and rod shaped., We report here the genome sequence of halophilic Halobacillus trueperi SS1, isolated from the Lunsu saltwater body in India. The bacteria are Gram positive and rod shaped. The genome of H. trueperi SS1 has 4.14 Mbp, with 4,329 coding sequences, 35 RNA genes (29 tRNAs, 2 rRNAs, and 4 noncoding RNAs), and 42.15% G+C content.

Sharma, K. D., Chen, W., and Muehlbauer, F. J. 2005. Genetics of chickpea resistance to five races of Fusarium wilt and a concise set of race differentials for Fusarium oxysporum f. sp. ciceris. Plant Dis. 89:385-390. Genetics of resistance in chickpea accession WR-315 to Fusarium wilt was investigated, and a concise set of differentials was developed to identify races of Fusarium oxysporum f. sp. ciceris. A population of 100 F7 recombinant inbred lines (RILs) from a cross of WR-315 (resistant) and C-104 (susceptible) was used to study genetics of resistance to races 1A, 2, 3, 4, and 5 of F. oxysporum f. sp. ciceris, and a population of 26 F2 plants from a cross between the same two parents was used to study inheritance of resistance to race 2. Segregations of the RILs for resistance to each of the five races suggest that single genes in WR-315 govern resistance to each of the five races. A 1:3 resistant to susceptible ratio in the F2 population indicated that resistance in WR-315 to race 2 was governed by a single recessive gene. A race-specific slow disease progress reaction was observed in chickpea line FLIP84-92C(3) to infection by race 2, a phenomenon termed as slow wilting, that is different from previously reported late wilting with respect to latent period, disease progress rate, and final disease rating. Twenty-nine germ plasm lines (27 Cicer arietinum and two C. reticulatum) including previously used differentials were evaluated for their reactions to infection by the five races. Only eight of the 29 germ plasm lines differentiated at least one of the five races based on either resistant or susceptible reactions, whereas the remaining germ plasm lines were either susceptible or resistant to all five races or differentiated them by intermediate reactions. A concise set of eight chickpea lines comprised of four genotypes and four F7 RILs with vertical resistance was developed as differentials for race identification in F. oxysporum f. sp. ciceris. These differential lines were characterized by early appearance of wilt symptoms, and clear and consistent disease phenotypes based on no wilt or 100% wilt incidence, which offers important improvements over previously available differential sets and provides more precise and unambiguous identification of the races.

Here, we report the genome sequence of hyperthermophilic and halophilic Parageobacillus toebii PW12, isolated from the Tattapani hot spring in the northwest Himalayas. The genome size of Parageobacillus toebii PW12 is 3,210,377 bp., Here, we report the genome sequence of hyperthermophilic and halophilic Parageobacillus toebii PW12, isolated from the Tattapani hot spring in the northwest Himalayas. The genome size of Parageobacillus toebii PW12 is 3,210,377 bp. The G+C content is 42.05%, and 3,382 coding sequences (CDS), 80 tRNAs, 5 noncoding RNAs (ncRNAs), and 4 CRISPR arrays were predicted.

We report here 16S rRNA-based bacterial diversity existing during freezing conditions in a high-altitude Himalayan lake through sequencing a 16S rRNA gene amplicon data set. A total of 121,857 high-quality reads were obtained; 40.78% of the bacterial population was classified to the genus level, while 1.26% was classified to the species level.

The internal transcribed spacer (ITS) region was used to discriminate 18 Saccharomyces cerevisiae strains isolated from fermented foods of the North-Western Himalayas. ITS sequences were compared with other known lineages available in literature. A phylogenetic tree grouped these strains into two clusters, the first comprising Malaysian, North American, Sake, West African and wine/European strains and the second comprising all Indian strains along with certain other strains from different countries. In both clusters, mixed grouping was observed, with the exception of a few strains with a region-based grouping pattern. The baking and brewing abilities of these strains were studied to expose functional diversity. Four strains, viz. Sc06, Sc11, Sc19 and Sc20, were found promising for baking and three strains, viz. Sc04, Sc05 and Sc24, were found promising for brewing, thereby indicating the existence of functional diversity. At a genetic level, mining of the ADH1 gene of ten randomly selected yeast strains, showed very little variation in the gene sequences irrespective of their alcohol production ability. In contrast, mining of the ATF1 gene showed a lot more variation in the gene sequences, revealing the ATF1 gene to be an appropriate marker to reveal differences in ester formation among these indigenous yeast strains.

Dactylorhiza hatagirea (D. Don) Soo, family Orchidaceae is an endangered medicinal herb inhabiting higher altitudes of western Himalayas. Due to reckless exploitation for its roots, it is of high conservation concern. In the present study fifteen microsatellites were developed and characterized across twenty collections of D. hatagirea. The total numbers of alleles amplified by these microsatellites were 64 with an average of 4.2 alleles per marker. Average observed and expected heterozygosity values for polymorphic loci were 0.623 and 0.631, respectively. Mean polymorphism information content value of the polymorphic markers was 0.532. Of the fourteen polymorphic microsatellites, 7 deviated from Hardy–Weinberg equilibrium. Microsatellites reported here can be utilized to address questions related to genetic characteristics in this species.

Translation initiation is down-regulated in eukaryotes by phosphorylation of the alpha-subunit of eIF2 (eukaryotic initiation factor 2), which inhibits its guanine nucleotide exchange factor, eIF2B. The N-terminal S1 domain of phosphorylated eIF2alpha interacts with a subcomplex of eIF2B formed by the three regulatory subunits alpha/GCN3, beta/GCD7, and delta/GCD2, blocking the GDP-GTP exchange activity of the catalytic epsilon-subunit of eIF2B. These regulatory subunits have related sequences and have sequences in common with many archaeal proteins, some of which are involved in methionine salvage and CO(2) fixation. Our sequence analyses however predicted that members of one phylogenetically distinct and coherent group of these archaeal proteins [designated aIF2Bs (archaeal initiation factor 2Bs)] are functional homologs of the alpha, beta, and delta subunits of eIF2B. Three of these proteins, from different archaea, have been shown to bind in vitro to the alpha-subunit of the archaeal aIF2 from the cognate archaeon. In one case, the aIF2B protein was shown further to bind to the S1 domain of the alpha-subunit of yeast eIF2 in vitro and to interact with eIF2Balpha/GCN3 in vivo in yeast. The aIF2B-eIF2alpha interaction was however independent of eIF2alpha phosphorylation. Mass spectrometry has identified several proteins that co-purify with aIF2B from Thermococcus kodakaraensis, and these include aIF2alpha, a sugar-phosphate nucleotidyltransferase with sequence similarity to eIF2Bvarepsilon, and several large-subunit (50S) ribosomal proteins. Based on this evidence that aIF2B has functions in common with eIF2B, the crystal structure established for an aIF2B was used to construct a model of the eIF2B regulatory subcomplex. In this model, the evolutionarily conserved regions and sites of regulatory mutations in the three eIF2B subunits in yeast are juxtaposed in one continuous binding surface for phosphorylated eIF2alpha.

Chickpea wilt caused by Fusarium oxysporum f. sp. ciceris is one of the major yield limiting factors in chickpea. The disease causes 10–90% yield losses annually in chickpea. Eight physiological races of the pathogen (0, 1A, 1B/C, 2, 3, 4, 5 and 6) are reported so far whereas additional races are suspected from India. The distribution pattern of these races in different parts of the world indicates regional specificity for their occurrence leading to the perception that F. oxysporum f. sp. ciceris evolved independently in different regions. Pathogen isolates also exhibit differences in disease symptoms. Races 0 and 1B/C cause yellowing syndrome whereas 1A, 2, 3, 4, 5 and 6 lead to wilting syndrome. Genetics of resistance to two races (1B/C and 6) is yet to be determined, however, for other races resistance is governed either by monogenes or oligogenes. The individual genes of oligogenic resistance mechanism delay onset of disease symptoms, a phenomenon called as late wilting. Slow wilting, i.e., slow development of disease after onset of disease symptoms also occurs in reaction to pathogen; however, its genetics are not known. Mapping of wilt resistance genes in chickpea is difficult because of minimal polymorphism; however, it has been facilitated to great extent by the development of sequence tagged microsatellite site (STMS) markers that have revealed significant interspecific and intraspecific polymorphism. Markers linked to six genes governing resistance to six races (0, 1A, 2, 3, 4 and 5) of the pathogen have been identified and their position on chickpea linkage maps elucidated. These genes lie in two separate clusters on two different chickpea linkage groups. While the gene for resistance to race 0 is situated on LG 5 of Winter et al. (Theoretical and Applied Genetics 101:1155–1163, 2000) those governing resistance to races 1A, 2, 3, 4 and 5 spanned a region of 8.2 cM on LG 2. The cluster of five resistance genes was further subdivided into two sub clusters of 2.8 cM and 2.0 cM, respectively. Map-based cloning can be used to isolate the six genes mapped so far; however, the region containing these genes needs additional markers to facilitate their isolation. Cloning of wilt resistance genes is desirable to study their evolution, mechanisms of resistance and their exploitation in wilt resistance breeding and wilt management.

Polo-like kinases (PLKs) belong to the serine/threonine kinase subfamily, characterized by the presence of the signature motif called Polo-box domain. PLK members studied so far have emerged as the conserved regulator of cell cycle and cell division in eukaryotes. The Polo-box domain adds diversity to PLK functions by targeting the enzyme to an array of substrates found at different sub-cellular structures for exquisite regulation of cell cycle. More than a dozen members of PLK subfamily have been identified in the eukaryotic world except in the higher plants. Despite the similarities in governing cell division, PLKs have diverse and unique functions in different organisms. This review summarizes the plethora of functions of PLK in yeast to humans. Along with its classical functions, this review also emphasizes on the role of PLKs in regulating DNA replication, repair, genome integrity, development, and morphogenesis pathways. Perturbations in PLK functions have disease implications, such as cancer in humans, and thus human PLK1 is targeted for cancer therapeutics. PLKs also play a vital role in regulating several stages of meiotic cell division. Thus, PLKs are emerging as a unique class of proteins with multiple and diverse functions in different organisms.

Sequence-tagged microsatellite site (STMS) and sequence-tagged site (STS) markers linked closely to Fusarium oxysporum f. sp. ciceris race 3 resistance gene in chickpea were identified, and linkage between three wilt resistance genes was elucidated. The resistance to race 3 in chickpea germplasm accession WR-315 was inherited as a single gene, designated foc-3, in 100 F(7) recombinant inbred lines derived from the cross of WR-315 (resistant) x C-104 (susceptible). The foc-3 gene was mapped 0.6 cM from STMS markers TA96 and TA27 and STS marker CS27A. Another STMS marker, TA194, at 14.3 cM, flanked the gene on the other side. Linkage between foc-3 and two other chickpea wilt resistance genes, foc-1 (syn. h(1)) and foc-4, was established. foc-3 was mapped 9.8 cM from foc-1 and 8.7 cM from foc-4, whereas foc-1 and foc-4 are closely linked at 1.1 cM. The identification of closely linked markers to resistance genes will facilitate marker-assisted selection for introgression of the race 3 resistance gene to susceptible chickpea lines.

In Neurospora crassa, multinucleate macroconidia are used for genetic transformation. The barrier for such a transformation can be either at the cell membrane level or at the nuclear membrane level. For assessment of these possibilities, a forced heterokaryon (containing two genetically marked nuclei and auxotrophic for histidine) of Neurospora crassa was transformed with a plasmid containing his-3(+) gene. The transformants, which could grow without histidine supplementation, were then resolved into component homokaryons to determine into which nucleus or nuclei the plasmid had entered. Our results suggest that the barrier for transformation in Neurospora crassa is at the nuclear level, not at the cell membrane level. In a heterokaryon containing two genetically distinct nuclei, plasmid DNA integrated into only one of the nuclear types at any instance, but never into both nuclear types. Thus, in Neurospora crassa, the competent nucleus is essential for the transformation event to take place, and at a given time only one type of nucleus is competent to take up the exogenous DNA. Genomic Southern analysis showed that the transformants harbor both ectopic and homologous integrations of the plasmid DNA. The type and number of integrations were reflected at the post-translational level, since the specific activity of histidinol dehydrogenase (the translation product of his-3+ gene) was variable among several transformants and always less than the level of the wild type.