苏云金芽胞杆菌4.0718菌株中杀虫晶体蛋白基因的定位及鉴定  

Localization and identification of crystal protein genes in Bacillus thuringiensis strain 4.0718

作　　者：付祖姣[1] 孙运军[1] 夏立秋[1] 丁学知[1] 胡胜标[1] 李文萍[1] 张友明[1] 

机构地区：[1]湖南师范大学生命科学学院,微生物分子生物学湖南省重点实验室,长沙410081

出　　处：《微生物学报》2008年第9期1250-1255,共6页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30670052);国家博士点基金(20060542006);国家“863计划”(2006AA02Z187,2006AA10A212)~~

摘　　要：【目的】对苏云金芽胞杆菌(Bacillus thuringiensis,简称Bt)4.0718菌株中的杀虫晶体蛋白基因(insecticidal crystal protein gene,简称cry基因)进行定位和鉴定,系统分析高毒力Bt4.0718菌株的杀虫基因背景。【方法】采用脉冲电泳(PFGE)分离Bt4.0718菌株的基因组DNA,确定该菌株的PFGE图谱和质粒图谱;采用Southern杂交分析该菌株中cry基因的定位,并使用PCR及PCR产物限制性片段长度多态性分析(PCR-RFLP)方法鉴定该菌株染色体和质粒上含有的cry基因类型。【结果】确定了Bt4.0718菌株的PFGE图谱和质粒图谱,鉴定到Bt4.0718菌株的染色体和质粒上均定位有cry基因,且分别包含cry1Aa、cry1Ac、cry2Aa和cry2Ab4种基因成分。但染色体上含有的cry基因可能不如质粒上含有的cry基因具有完整的开放阅读框。【结论】首次在Bt4.0718菌株的染色体上发现有丰富的cry基因,且与质粒上含有的cry基因类型一致。[Objective] To localize and identify the insecticidal crystal protein genes （cry genes） in high-toxic Bacillus thuringiensis strain 4.0718. [Methods] The genomic DNA of B. thuringiensis strain 4.0718 was isolated by pulse field gel electrophoresis （PFGE）, and the PFGE pattern and plasmid pattern were characterized. The cry genes in the strain were analyzed by Southern blot and the cry gene types on the chromosome and plasmid were identified by PCR and restriction fragment length polymorphism patterns of the PCR-amplified fragments （PCR-RFLP）. [Results] Southern blots showed that the cry genes in B. thuringiensis strain 4.0718 are present on both chromosome and plasmid. PCR and RFLP patterns showed that the chromosome and plasmid contain four cry genes, crylAa, crylAc, cry2Aa, and cry2Ab. However, the cry genes on the chromosome may be incomplete, while the crylAa and crylAc genes localized on the plasmid were found to contain the full coding sequence. [Conclusion] The abundant cry genes were first found on chromosome of B. thuringiensis strain 4.0718, and the cry-type genes are the same as the plasmid.

关 键 词：苏云金芽胞杆菌 CRY基因 定位 脉冲电泳(PFGE) 限制性片段长度多态性分析(RFLP) 

分 类 号：S476[农业科学—农业昆虫与害虫防治]