摘要
利用合成的寡聚核苷酸引物及 PCR 扩增技术对 B.t.k-δ-内毒素基因进行了克隆和修饰。在大肠杆菌中表达的抗虫试验表明该修饰过的 B.t.k-δ-内毒素基因对鳞翅目昆虫如烟青虫和棉铃虫有较强的毒杀作用。随后,通过农杆菌介导将该基因转入烟草,RNA 点杂交及抗虫试验表明 Bt 基因在烟草中得到表达,其表达水平足以防治烟青虫的为害。
B.t.k.-δ-endotoxin gene(abbreviated:Bt gene)was cloned and modified by using synthetic oligonucleotide primers and PCR amplification method.The Bt gene was first expressed in E.coli,insect boiassays of the total bacterial protein proved that the Bt gene product was lethal both to tobacco horn- worm(Manduca sexta)and cotton bollworm(Heliothis zea).Then,the Bt gene was transferred into to- bacco plants by leaf disco method,expression of the gene in tobacco plants provided effective control of Manduca sexta.
关键词
内毒素
大肠杆菌
基因表达
抗虫性
烟草
B.t.k.-δ-endotoxin
gene cloning
gene expression
insect-resistance
