抗虫基因 Cry1C*重组表达载体的构建与鉴定

摘要

To provide strains for biological control of rice pests,the insect-resistant gene Cry 1C from GenBank was optimized,added BamH I and Spe I restriction enzyme cutting sites at both sides,and artificially synthesized.The modified Bt gene was named Cry 1C * ,which was inserted into pTCK303 after double restriction-enzyme digestion to construct recombinant expression vector pTCK303-Cry 1C * ,and transferred into A.tumefaciens LBA4404.The PCR,double digestion,and sequencing results showed that the recombinant vector was successfully constructed(2 342 bp).The PCR result showed that the expression vector pTCK303-Cry 1C * had been transformed into A.tumefaciens LBA4404,successfully constructing a genetic engineering bacteria.%为水稻害虫生物防治提供菌株，从GenBank中选定Cry 1C 类抗虫基因，将其优化改造后命名为 Cry 1C *，Cry 1C *经 BamH I 和 Spe I 双酶切后与植物表达载体 pTCK303连接，构建 pTCK303-Cry 1C *重组表达载体，将其转化到农杆菌 LBA4404中，同时进行 PCR、双酶切及测序鉴定。结果表明：PCR、双酶切及测序产物与预期扩增片段（2342 bp）相符，重组表达载体 pTCK303-Cry 1C *构建成功；农杆菌LBA4404中含有重组质粒，成功构建了 Cry 1C *农杆菌基因工程菌株。