Bt4.0718 cry1Ac基因的克隆与表达分析(英文)  

Cloning and Expression Analysis of the cry1Ac Gene from Bacillus thuringiensis Strain 4.0718

作　　者：丁学知[1] 张何[1] 孙运军[1] 黄潢[1] 张春艳[1] 夏立秋[1] 

机构地区：[1]湖南师范大学生命科学学院,湖南长沙410081

出　　处：《激光生物学报》2006年第6期563-570,共8页Acta Laser Biology Sinica

基　　金：The National Natural Science Foundation of China(No.670052);Scientifical Item of Education Depart-ment of Hunan Province(No.02C217)and the Natural Science Foundation of Hunan Province(No.06JJ2009).

摘　　要：在基因库中比对14种cry1Ac基因序列,发现了同源性很高的上游启动子区域和下游终止子区域。根据这一同源序列设计引物,从B t4.0718中扩增出包含双启动子和终止子的4.2 kb片段,用PCR-RFLP检测确定其中含有cry1Ac基因。然后将此片段克隆到穿梭载体pHT304中,转化大肠杆菌DH5α和B t无晶体突变株XZM-101。同时,利用原子力显微镜观察发现重组菌株BXZM34能够产生菱形晶体。14 cry1Ac genes from GenBank were aligned and the consensus regions in the upstream of promoter and in the downstream of terminator were found. Based on the consensus sequences, a pair of primers was designed and a 4.2 kb element was amplified that includes the dual overlapping promoter and the whole termination-associated sequence from Bacillus thuringiensis strain 4.0718, and the amplified 4.2 kb element was confirmed to contain the cry1Ac gene by cryl sub-genetype PCR-RFLP cry gene typing system. The 4. 2 kb element was cloned into Bt-E. coli shuttle vector pHT3.04 and the cry1Ac gene was also expressed in E. coli DHSα and acrystalliferous mutant XZM-101. Meanwhile, rhombic crystal was observed from recombinant strain BXZM34 by atomic force microscope.

关 键 词：苏云金芽孢杆菌 CRY1AC基因 穿梭载体 重组菌株 

分 类 号：Q785[生物学—分子生物学] Q786