苏云金芽胞杆菌 LSZ 9408菌株基因型的鉴定

基金项目:

国家"948"项目子专题(2011-G25);福建省科技计划项目——省属公益类科研院所基本科研专项(2009R10037-1);福建省杰出青年科学基金项目(2009J06010)

Identification of Cry-type Gene of Bacillus thuringiensis Strain LSZ 9408

1.
Agricultural Bio-Resources Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350003, China

摘要: 利用聚合酶链式反应(PCR)和聚丙烯酰胺凝胶电泳(SDS-PAGE)技术鉴定苏云金芽胞杆菌 LSZ 9408菌株的杀虫晶体蛋白及其基因组成。PCR结果表明:苏云金芽胞杆菌 LSZ 9408菌株中含有cry1和cry2两种基因型。采用5对通用引物—Un1、Un2、Un3、Un4、Un7/8进行PCR扩增,结果显示:Un1和Un2引物扩增的DNA 片段大小分别为270 bp和700 bp左右,其他引物无PCR扩增产物。SDS-PAGE结果也表明:苏云金芽胞杆菌 LSZ 9408菌株中含有cry1和cry2两种基因,它们编码的杀虫晶体蛋白分子量约为130 kD和65 kD。
- LSZ 9408菌株 /
- 杀虫晶体蛋白基因 /
- 聚合酶链式反应 /
- 聚丙烯酰胺凝胶电泳
Abstract: PCR and SDS-PAGE were used to identify the cry-type gene of Bacillus thuringiensis (Bt) strain LSZ 9408. PCR analysis showed that Bt LSZ 9408 strain contained cry1 and cry2 genes. Five pairs of general primers-Un1, Un2, Un3, Un4 and Un7/8 were used to amplify PCR, and then the amplification results were detected by SDS-PAGE. Primes Un1 and Un2 formed about 270 bp and 700 bp DNA fragments on agarose gel electrophoresis, while none PCR products was found with the other three pairs of primers. The results of SDS-PAGE also indicated that Bt LSZ 9408 possessed cry1 and cry2 genes, which coded 130kD and 65kD ICPs.
- Bt LSZ 9408 strain /
- cry-type gene /
- PCR analysis /
- SDS-PAGE

[1]	冯纪年, 李陇梅, 张兴. 苏云金芽胞杆菌新菌株S249 cry2Ad2的克隆及特征分析[J].西北农林科技大学学报, 2006, 34(5): 93-96.
[2]	CAROZZI N B,DRAMER V C,WALLEN G W. Prediction of insecticidal activity of Bacillus thuringiensis strains by polymerase chain reaction product profiles[J]. Appl Environ Microbiol,1991, 57: 3057-3061.
[3]	CERON J. PCR analysis of the cry1 insecticidal crystal family genes from Bacillus thuringiensis[J]. Appl Environ Microbiol,1994, 60: 353-356.
[4]	郭艾英, 王硕.苏云金芽胞杆菌晶体蛋白的制备方法[J]. 食品与发酵工业, 2005, 31(8): 8-10.
[5]	左雅慧, 丁之铨, 张杰.苏云金芽胞杆菌培养条件及晶体蛋白提纯方法初探[J]. 植物保护, 1999, 25(4): 32-34.