绿脓杆菌外毒素A受体结合区蛋白基因重组与鉴定  

Recombination and Identification of PEA Receptor Binding Subunit Gene

作　　者：万忠海[1] 王景林[1] 江禹[1] 刘晓明[1] 何畅[1] 朱平[1] 

机构地区：[1]解放军军需大学军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2002年第6期571-573,共3页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目 ( 39770 86 6 )

摘　　要：为了获得高表达量重组绿脓杆菌外毒素 A(PEA)受体结合区蛋白 ,以用作免疫原 ,从含有目的片段 (10 0 0 bp)的 p ET- 2 0 (b) +B1 0 质粒切下目的片段 ,以正确阅读框架插入含有 T7lac启动子的 p ET- 2 8c(+) DNA中 ,构建了p ET- 2 8c(+) B1 0 质粒。用该质粒转化大肠杆菌 BL2 1 (DE3 )、BL2 1 (DE3 ) p L y S,经 IPTG诱导 ,BL2 1 (DE3 ) p L y S能高效表达目的蛋白。 SDS- PAGE、Western blot分析证实 ,该蛋白即是所需的蛋白 ,该重组蛋白占菌体总蛋白的 5 8%。For the purpose of obtaining a high level expression of recombinant protein of the receptor binding domain of pseudomonas exotoxin A (PEA),the vector pET-28c(+)B 10 that can express the peptide was constructed.The target gene-B 10 was cut out from pET-20(b)+B 10,then it was inserted in pET-28c(+) DNA just follow the T 7 lac promotor.The recombinant plasmid pET-28c(+)B 10 was transmitted into E.coli BL 21(DE 3) and BL 21(DE 3)pLyS.The strains BL 21(DE 3)pLyS can express the target protein at high level.By SDS-PAGE and Western-blot the protein was proved to be the target protein,it accounts for 58% of the total amount of the bacteria protein.

关 键 词：绿脓杆菌外毒素A 受体结合区 基因重组 鉴定 

分 类 号：S852.61[农业科学—基础兽医学]