苏云金芽孢杆菌Rpp02菌株cry1 Ac20基因的克隆及表达(英文)  

Cloning and expression of cry1Ac20 gene from Bacillus thuringiensis strain Rpp02

作　　者：谭芙蓉[1] 李平[1] 郑爱萍[2] 周华强[1] 郑秀丽[1] 

机构地区：[1]四川农业大学水稻研究所 [2]四川省农业生物技术工程中心

出　　处：《昆虫学报》2006年第6期950-954,共5页Acta Entomologica Sinica

基　　金：国家"863"高技术研究发展计划项目(2002AA212151);教育部长江学者和创新团队发展计划(IRT0453)

摘　　要：Rpp02菌株是本实验室分离的一株对鳞翅目等多种害虫具有高毒力的苏云金芽孢杆菌莫里逊亚种(Bacillus thuringiensis subsp.morrisoni),经PCR检测,它含有cry1Ac基因。对其基因组DNA进行PCR扩增,得到大约4kb的产物。测序结果表明,该片段含有一个较大的ORF框,基因编码区为3534bp,编码1177个氨基酸,分子量为133.144kD,pI4.952,为弱酸性蛋白质,亮氨酸(Leu)、丝氨酸(Ser)、谷氨酸(Glu)3种氨基酸含量最高,分别为8.0%、7.8%、7.7%。该基因序列与cry1Ac序列同源性达到99%,并被国际Bt杀虫晶体蛋白基因命名委员会命名为cry1Ac20。生物测定表明,该基因在大肠杆菌中得到了表达,表达产物具有较强的杀虫效果,试喂菜青虫48h后,校正死亡率为88.78%。Bacillus thuringiensis subsp, morrisoni strain Rpp02 isolated by researchers in our laboratory was highly toxic to several kinds of insect posts. Results of PCR analysis indicated that strain Rpp02 contained the cry 1 Ac gene, and subsequently a novel cry l Ac gene was cloned from this strain by PCR techniques. The results of sequence analysis showed that the novel gene crylAc20, named by B. thuringiensis Pesticidal Crystal Protein Nomenclature Committee, contained an open reading frame of 3 534 nucleotides encoding a protein of 1 177 amino acids with a predicted molecular mass of 133. 144 kDa and isoelectric point of 4.952. Compared with other known cry 1 Ac genes, cryl Ac20 has shown as high as 99% nucleotide sequence homology. Bioassay showed that the toxic protein appeared high insecticidal activity against Pieris rapae L., and the corrected mortality was 88.78 % 48 h after feeding with leaves immersed in cultures containing transformants.

关 键 词：苏云金芽孢杆菌 cry1Ac20基因 克隆 表达 菜青虫 

分 类 号：Q965[生物学—昆虫学]