J Immunol Methods. https://doi.org/10.1371/journal.pone.0139822.g001. https://doi.org/10.1371/journal.pone.0139822.s001. During housing, all animals were monitored twice daily to assess their health status. The samples are described in Table 2. This study was approved by the Ethical Committee of Animal Experiments of the College of Veterinary Medicine at Nanjing Agricultural University. | The PCR products were loaded onto 8% (wt/vol) polyacrylamide (37:1 acrylamide/bisacrylamide) gels in 1× TAE buffer with a denaturing gradient ranging from 30 to 60%. -, Theriogenology. Yes Ethics statement . 2013ZX08008-004 and 2014ZX08008-004) and A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions. Yes No GH and neoR were detected in the intestinal content, feces and soil samples, suggesting that no horizontal gene transfer occurred in the course of this study. National Center for Biotechnology Information, Unable to load your collection due to an error, Unable to load your delegates due to an error. The microbial community of the gastrointestinal tract is closely associated with the host metabolism and has a complex and sensitive construction [11]. Yes All the fresh samples are stored in -70°C before analysis. Bootstrap confidence levels greater than 50% are indicated at the nodes (replicate 1,000 times). Feces were taken from GH transgenic and non-transgenic goats, and each sample was taken when it just been defecated. The 16S rDNA genes were amplified with an initial denaturation of DNA at 94°C for 10 min, followed by 30 cycles of 60 s of denaturation at 94°C, 60 s annealing at 55°C, 90 s of elongation at 72°C, and a final elongation for 10 min at the same temperature. Female GH transgenic and non-transgenic Saanen dairy goats were raised on a farm in the Transgenic Research Center, Shanghai, China. 1997) and the feasibility of the large-scale production for the industrial application has been confirmed (Baldassarre et al. All animal care and use procedures were conducted in strict accordance with the Animal Research Committee guidelines of the College of Veterinary Medicine at Nanjing Agricultural University. Moura RR, Lopes-Junior ES, Teixeira DI, Serova IA, Andreeva LE, Melo LM, Freitas VJ. 1999 May;17(5):456-61 Production of transgenic founder goats involves introducing and stably integrating an engineered piece of DNA into the genome of the animal. We minimized the risk of horizontal gene transfer in the GE goats, although it still could occur in the gut and rumen. broad scope, and wide readership – a perfect fit for your research every time. Substantially contributed to the concept and design of the work: QY QHY. The results showed that the extracted DNA from all samples contained the bacterial gene. here. here. PLOS ONE promises fair, rigorous peer review, 2019 Jan;12(1):12-33. doi: 10.14202/vetworld.2019.12-33. Scale bars indicate differences among the profiles. Any changes in the gastrointestinal bacterial flora could thus conceivably also influence the surrounding environmental soil flora [12]. Yes Epub 2016 Apr 21. 2016 Jul 1;86(1):160-9. doi: 10.1016/j.theriogenology.2016.04.028. https://doi.org/10.1371/journal.pone.0139822.g003. COVID-19 is an emerging, rapidly evolving situation. Over the time period indicated (1995-2012), pronuclear microinjection was used in a number of programs to insert transgenes into 18,120, 1- or 2- cell stage fertilized embryos. Then, these recombinant plasmids were transformed into Escherichia coli DH5α. PC: positive control with pcGH vector over-expression. Numerous monoclonal antibodies are currently being produced in the mammary glands of transgenic goats [9] and cloned transgenic cattle have been created, which produce recombinant bispecific antibody in their blood. Seven groups could be found based on the phylogenetic distribution of the 16S rDNA cloning libraries: Firmicutes, Bacteroidetes, Proteobacteria, Acitinobacteria, Chloroflexi, Nitrospirae and Acidobacteria. The development of genetically engineered animals has brought with it increasing concerns about biosafety issues. Three or five clones were randomly selected for each band. We also demonstrated that the foreign GH gene and neoR gene did not change the microbial flora in the goat intestine or the surrounding soil. All the goats were healthy and fed with the same fodder (Table 1). Analyzed the data: ZB XG QZ ZB XG. The breeding of animals using genetically engineered (GE) technology has recently become possible. Gavin W(1), Blash S(1), Buzzell N(2), Pollock D(1), Chen L(1), Hawkins N(1), Howe J(1), Miner K(1), Pollock J(1), Porter C(1), Schofield M(1), Echelard Y(1), Meade H(1). To study the influence of the GH and neoR genes on the bacterial community structure, DGGE was performed after PCR amplification of the variable V3 region of the 16S rDNA from the microbial DNAs. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China, Affiliation Yes The band patterns for the feces, soil and intestinal content samples showed degrees of similarity that were higher than 96%, 96.5% and 95%, respectively (Fig 3). Find NCBI SARS-CoV-2 literature, sequence, and clinical content: https://www.ncbi.nlm.nih.gov/sars-cov-2/. No, Is the Subject Area "Denaturing gradient gel electrophoresis" applicable to this article? Horizontal gene transfer, also known as lateral gene transfer, refers to the transfer of genes between different species, such as between prokaryotes and eukaryotes in a manner other than traditional reproduction [6]. HHS (2015) The Effects of GH Transgenic Goats on the Microflora of the Intestine, Feces and Surrounding Soil. We slaughtered the goats and cut the intestine lengthwise to collect the intestinal contents from jejunum and cecum. No adverse events were observed. Performed the experiments: ZB XG QZ JL WH HQY JC. -, Biotechnology (N Y). Growth hormone; SCNT, No, Is the Subject Area "Polymerase chain reaction" applicable to this article? Affiliation The numbers in the figures indicate the DGGE bands selected for cloning and sequencing. -. This work was supported by grants from the National Animal Transgenic Breeding Grant Project (No. Her name comes from a protein she carries in her milk, glucocerebrosidase, which works to process glucocerebrosides, a type of cell lipid.
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