利用流式细胞仪分选土壤中发生自然转化的阳性转化子

发布时间：2018-05-13 09:10

  本文选题：水平基因转移 + 自然转化　； 参考：《华中农业大学学报》2016年02期

【摘要】：为确定土壤中的DNA通过自然转化的方式发生水平基因转移的频率,将1个带有egfp和卡那霉素抗性基因Kanr的质粒DNA添加到自然土壤中,分离菌悬液,上流式细胞仪分选能发出绿色荧光且具有卡那霉素抗性的阳性转化子,确定土壤中发生自然转化的频率小于10~(-7)。分离的菌悬液通过培养基的富集,从39 736 426个细胞中分选获得了82个细胞,最终得到了纯化的阳性转化子,证实土壤中的DNA通过自然转化发生了水平基因转移。
[Abstract]:In order to determine the frequency of horizontal gene transfer of DNA in soil by natural transformation, a plasmid DNA containing egfp and kanamycin resistant gene Kanr was added to natural soil to isolate bacterial suspension. The up-flow cytometer was used to separate the positive transformants with green fluorescence and kanamycin resistance, and the frequency of natural transformation in soil was determined to be less than 10 ~ (10) ~ (-7) ~ (th). 82 cells were isolated from 39 736 426 cells and purified positive transformants were obtained, which confirmed that horizontal gene transfer took place through natural transformation of DNA in soil.
【作者单位】： 华中农业大学农业微生物学国家重点实验室;
【基金】：国家自然科学基金项目(41071180,41230854) 国家“863”计划项目(2012AA101402)
【分类号】：S154
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本文编号：1882535