中国畜牧兽医2012，Vol.39Issue(12)：10-13,4.
D型产气荚膜梭菌α毒素基因的克隆与高效表达
Cloning and Expression of α Toxin Gene from Toxin Type D Clostridium perfringens
摘要
Abstract
To establish the method for cloning and expression of Clostridium perfringens a toxin gene(cpa) ,the cpa gene was amplified by PCR from toxin type D strain,then was inserted into pET-28b to construct pET-28b-cpa. Identified by PCR, restriction enzyme digestion and sequencing methods,the recombinant plasmid was transformed into BL21 (DE3) pLysS and induced to express by IPTG. The size and distribution of the target protein were detected by SDS-PAGE,and its reactionogenicity was confirmed by Western blotting. The results showed that the cpa gene was 1110 bp and the homology with reference sequence of GenBank was greater than 99%. In SDS-PAGE analysis,the target protein was 41. 2 ku as expected and distributed in ultrasonic lysis supernatant as well as in inclusion bodies,but mainly existed in inclusion bodies. Both of them showed similar reactionogenicity to native a toxin.关键词
产气荚膜梭菌/α毒素/克隆/表达Key words
Clostridium perfringens /α toxin/cloning/expression分类
农业科技引用本文复制引用
李娜,吴建勇,李建军,杨学云,王登峰,段新华,王治才..D型产气荚膜梭菌α毒素基因的克隆与高效表达[J].中国畜牧兽医,2012,39(12):10-13,4.基金项目
农业公益性行业科研专项(201103008) （201103008）
新疆多胎(多羔)肉羊杂交生产体系建设行动计划项目. （多羔）
