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苏云金杆菌4.0718质粒上杀虫晶体蛋白基因的PCR分析
引用本文:夏立秋,张何,刘全兰,陈宇,莫湘涛.苏云金杆菌4.0718质粒上杀虫晶体蛋白基因的PCR分析[J].生物技术通报,2002(2):35-37,46.
作者姓名:夏立秋  张何  刘全兰  陈宇  莫湘涛
作者单位:湖南师范大学生命科学学院微生物学系,长沙,410081
基金项目:湖南省青年科技基金项目 ( 96 -32 -0 9),湖南省农业重大攻关项目 ( 99-10 0 4-0 8)资助
摘    要:苏云金杆菌(Bacillus Thuringiensis)4.0718由本实验室选育是对鳞翅目(Lepidopteran)昆虫有高毒性的菌株。采用SDS温和提取方法提取此菌株的质粒,电泳纯化后获得了其4种不同分子量的质粒P1,P2,P3,P4。利用序列分析软件对已知的cryⅠ类型基因进行阵列比较,根据其高度保过区域设计了1对通用引物,对cryⅠ类型基因的扩增产物对277bp左右的片段,用此引的对Bacillus thuringiensis 4.0718的4种不同质粒进行了PCR分析,发现质粒P1,P2,P3扩增阳性,都产生了277bp的扩增片段,而质粒P4没有扩增产物,这为进一步的基因定位打下了基础。

关 键 词:苏云金杆菌4.0718  质粒  杀虫晶体蛋白  PCR分析  cryⅠ基因

PCR- based Analysis of Insecticidal Crystal Proteins Genes on Plasmids from Bacillus Thuringiensis 4.0718
Xia Liqiu,Zhang He,Liu Quanlan,Chen Yu,Mo Xiangtao.PCR- based Analysis of Insecticidal Crystal Proteins Genes on Plasmids from Bacillus Thuringiensis 4.0718[J].Biotechnology Bulletin,2002(2):35-37,46.
Authors:Xia Liqiu  Zhang He  Liu Quanlan  Chen Yu  Mo Xiangtao
Abstract:Bacillus Thuringiensis strain 4.0718 is highly toxic to lepidopteran.Plasmids are extracted from this strain by using SDS gentle m,ethod .According to the result of agarose gel electrophoresis,we know this strain comprises four kinds of different plasmids P1?P2?P3 and P4. Perhaps some include cryI type genes.One pair of universal primers for cryI genes was designed to amplify a 277 bp fragment by simultaneous alignment with all previously described cryI genes.The 277 bp predicted fragment can be amplified from P1?P2 and P3,but P4 is negative to the primers,which indicates P1 and P2 and P3 all include cryI type genes,but P4 does not. These resuts can help us find the useful genes for the gene engineering.
Keywords:Bacillus thuringiensis  PCR  CryI gene
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