中文摘要：新霉素磷酸转移酶（neo）基因被广泛用为基因工程动植物生产中的选择性标记。neo转基因表达的安全性问题近期被广泛关注。本文将neo转基因仔猪和非转基因仔猪随机分为A、两组，并使用高通量测序技术对肠道菌群的变化进行研究，来评价neo基因的影响。其中，A组猪喂养标准饮食并添加抗生素新霉素；B组猪喂养标准饮食。本文用多重PCR技术研究外源新霉素磷酸转移酶基因的水平转移；并通过监测A组动物肠道菌群变化的不同模式发现，若转基因猪小肠中分泌NPTⅡ可防护新霉素。研究发现，从转基因组织/细胞向肠道菌群分泌NPTⅡ会导致新霉素活性受抑制以及细菌数量不同程度的变化；neo基因也会对标准喂养（未增加新霉素）的转基因猪的肠道菌群产生意想不到的效果。研究结果表明，应仔细考虑和充分评估neo转基因在基因工程动物的广泛应用问题。
外文摘要：The neo (neomycin phosphotransferase) gene is widely used as a selection marker in the production of genetically engineered animals and plants. Recent attention has been focused on safety concerns regarding neo transgene expression. In this study, neo transgenic and non-transgenic piglets were randomly assigned into Group A and Group B to evaluate effects of neo transgene by studying changes in gut microbiota using high-throughput sequencing. Group A pigs were fed a standard diet supplemented with antibiotic neomycin; Group B pigs were fed a standard diet. We examined horizontal transfer of exogenous neo gene using multiplex PCR; and investigated if the presence of secreted NPT II (neo expression product) in the intestine could lead to some protection against neomycin in transgenic pigs by monitoring different patterns of changes in gut microbiota in Group A animals. The unintended effects of neo transgene on gut microbiota were studied in Group B animals. Horizontal gene transfer was not detected in gut microbiota of any transgenic pigs. In Group A, a significant difference was observed between transgenic pigs and non-transgenic pigs in pattern of changes in Proteobacteria populations in fecal samples during and post neomycin feeding. In Group B, there were significant differences in the relative abundance of phyla Firmicutes, Bacteroidetes and Proteobacteria, and genera Lactobacillus and Escherichia-Shigella-Hafnia between transgenic pigs and non-transgenic pigs. We speculate that the secretion of NPT II from transgenic tissues/cells into gut microbiota results in the inhibition of neomycin activity and the different patterns of changes in bacterial populations. Furthermore, the neo gene also leads to unintended effects on gut microbiota in transgenic pigs that were fed with basic diet (not supplemented with neomycin). Thus, our data in this study caution that wide use of the neo transgene in genetically engineered animals should be carefully considered and fully assessed.
作者：Qingqing Wang；Lili Qian；Shengwang Jiang；Chunbo Cai；Dezun Ma；Pengfei Gao；Hegang Li；Ke Jiang；Maoxue Tang；Jian Hou；Jie Liu；Wentao Cui
作者单位：中国农业科学院
期刊名称：PLOS ONE
期刊影响因子：3.057
出版年份：2016
出版刊次：3
点击下载：通过基于高通量测序技术地肠道菌群变化研究来评价Neo转基因猪的安全性