摘要
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(Bacillus atrophaeus)ATCC 9372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,ATCC 9372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-Hind III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Yth A基因和Yti B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ATCC 9372-Yti B的Cr(VI)去除率下降了约12%,表明Yti B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。
In order to verify the feasibility of PFGE technology on finding out the different genes associated with Cr（ VI）removal between ATCC 9372 and Ua strains,the strains were cultured by LB medium which contains 0,20,40,60,80mg·L^-1Cr（ VI）,OD600 and removal rates were measured at 0,12,24,36,48,60 h. PFGE technology was used to study the differences of DNA between two strains which have been enzyme. The feasibility of PFGE technology was verified by PCR and analysis of restriction sites. GM technology was used to test the function of different genes on removing Cr（ VI）. Results showed that the growth conditions and removal rates of ATCC 9372 and Ua strains reached the maximum at 48 h,ATCC 9372 strain grew better than Ua strain,and its removal rate reached 95% in 40 mg·L^-1Cr（ VI） solution as well as the removal rate of Ua strain just reached 52. 31%. There is a 3Kb different fragment which had been digested by EcoRI-Hind III between ATCC9372 and Ua strains was showed by the results of PFGE technology.Yth A and Yti B genes were contained in the different fragment. The feasibility of PFGE to explore difference among DNA is credible which was tested by PCR and the analysis of restriction sites. The removal rate of genetically modified strain ATCC 9372-Yti B declined 12% reflected that gene Yti B can inhibit the function of removing Cr（ VI）. Our study provide a new approach of searching different genes theoretically,and the approach need further improvement.
出处
《核农学报》
CAS
CSCD
北大核心
2015年第6期1052-1060,共9页
Journal of Nuclear Agricultural Sciences
基金
国防科技项目(14zg6101)
国家"973"计划(2014CB846003)
四川省生物质资源利用与改性工程中心项目(13zxsk02)
关键词
铬污染
萎缩芽孢杆菌
脉冲场凝胶电泳
基因差异分析
Yti
B基因
chromium pollution Bacillus atrophaeus pulsed field gel electrophoresis genetic differences analyze gene YtiB
