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核农学报  2015 

2株萎缩芽孢杆菌铬去除差异基因的PFGE分析

DOI: 10.11869/j.issn.100-8551.2015.05.1052, PP. 1052-1060

Keywords: 铬污染,萎缩芽孢杆菌,脉冲场凝胶电泳,基因差异分析,YtiB基因

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Abstract:

为寻找2株萎缩芽孢杆菌与铬去除相关的差异基因,探索PFGE技术在寻找差异基因方面的可行性,本研究以萎缩芽孢杆菌(Bacillusatrophaeus)ATCC9372菌株和Ua菌株为材料,分别考察2株菌对Cr(VI)的耐受性和去除能力;采用PFGE技术进行基因组酶切片段分析,以期获得差异基因;利用转基因技术对得到的差异基因Cr(VI)去除功能进行验证。结果表明:2株菌在48h时菌体生长和对Cr(VI)的去除率达到最大值,ATCC9372菌株的生长状况和Cr(VI)去除能力明显高于Ua菌株;基因组通过内切酶EcoRI-HindIII组合酶切后,PFGE分析得到了一条3kb左右的差异条带,测序比对结果表明该序列含有YthA基因和YtiB基因;差异序列的酶切位点与内切酶组合一致,PCR结果表明差异序列只存在于Ua菌株中,转基因菌株ATCC9372-YtiB的Cr(VI)去除率下降了约12%,表明YtiB基因与Cr(VI)的去除负相关。本研究利用PFGE技术成功发现2株萎缩芽孢杆菌的铬去除差异基因,经验证该技术可行性高,为寻找差异基因提供了一种新途径。

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