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苏云金芽孢杆菌4.0718菌株的杀虫晶体蛋白基因分析
引用本文:丁学知,刘全兰,莫湘涛,高必达,夏立秋.苏云金芽孢杆菌4.0718菌株的杀虫晶体蛋白基因分析[J].微生物学报,2003,43(3):413-417.
作者姓名:丁学知  刘全兰  莫湘涛  高必达  夏立秋
作者单位:1. 湖南师范大学生命科学学院微生物学系,长沙,410081;湖南农业大学植物保护学院,长沙,410128
2. 湖南师范大学生命科学学院微生物学系,长沙,410081
3. 湖南农业大学植物保护学院,长沙,410128
基金项目:国家自然科学基金项目 (30 2 70 0 37),国家“863计划”项目 (2 0 0 2AA2 4 50 2 1),华中农业大学农业部农业微生物重点实验室项目 (3- 0 2 - 1- 2 1)~~
摘    要:根据苏云金杆菌(Bacillus thuringiensis)cry1、cry2和cry3型基因的保守区分别设计了3对通用引物Un1(d)/Un1?、Un2(d)/Un2?和Un3(d)/Un3?,以Bt4.0718菌株质粒DNA为模板进行PCR扩增,通过扩增产物片段的分子量大小来确定该菌株所含有的杀虫晶体蛋白基因类型。随后根据上述3类cry基因的高变区设计特异引物再次进行PCR鉴定。结果表明:Bt4.0718菌株含有cry1Aa、cry1Ab、cry1Ac、cry1Cb、cry2Ac和新基因cry4.5等6种基因类型。这一结果为利用该菌株构建高效广谱杀虫工程菌提供了客观依据。

关 键 词:苏云金芽孢杆菌,杀虫晶体蛋白基因,PCR
文章编号:0001-6209(2003)03-0413-05

Characterization of Insecticidal Crystal Proteins Genes from Bacillus thuringiensis 4.0718 Strain
Ding Xuezhi , Liu Quanlan Mo Xiangtao Gao Bida Xia Liqiu.Characterization of Insecticidal Crystal Proteins Genes from Bacillus thuringiensis 4.0718 Strain[J].Acta Microbiologica Sinica,2003,43(3):413-417.
Authors:Ding Xuezhi  Liu Quanlan Mo Xiangtao Gao Bida Xia Liqiu
Institution:Department of Microbiology, College of Life Sciences, Human Normal University, Changsha 410081, China.
Abstract:In this study, we rapidly identified Bacillus thuringiensis 4.0718 strain that harbored the known cry1 and cry2 type genes by a PCR strategy. Three pairs of universal oligonucleotide primers were designed to detect all known cry1, cry2 and cry3 type gene sequences. Then the DNA of the positive strain 4.0718 was probed with a set of specific primers. One feacture of this screening method was that each gene was expected to produce a PCR product having a precise molecular weight. PCR products having different sizes probably represented the gene was a potentially novel gene. Differentiations among these genes was determined on the basis of the electrophoresis patterns of PCR products. Finally, five cry1 type genes (cry1Aa, cry1Ab, cry1Ac, cry1Cb, a novel cry4.5 type genes) and one cry2Ac type gene had been detected from Bacillus thuringiensis 4.0718 strain.
Keywords:Bacillus thuringiensis  Insecticidal crystal protein gene  PCR
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