摘要
用多聚酶链反应(PCR)和基因重组技术通过百日咳毒素S_3亚基基因和高表达质粒pMAL-C构建了麦芽糖结合蛋白(MBP)和(S_3)亚基蛋白的融合基因,并在大肠杆菌中高效表达了(MBP-S_3)融合蛋白。该融合蛋白产率占细胞总蛋白含量的25%,并且能用蛋白酶因子Xa将融会蛋白切开,得到MBP(42KD)和S_3亚基蛋白(21.87KD).该方法表达式效率高。纯化过程简单,为基因工程技术生产该毒素提供了新的途径。
sing Polyinerase Chain Reaction(PCR)and recombinant DNA techniques,a fusing genevector inc luding maltose binding protein(MBP)and pertussis toxin S3(PTX-S3) subunit proteinis constructed by PTX-S3 gene and a high level expressioa plasmid pMAL-C. High levelexpressi on of the fusing protein may be produced by the fusing pla smid in E. coli,the fusingprotein (MW,63.87KD)yield is about 25% in the whole cell protein. MBP(MW,42KD) andPTX-S3(MW,21.87KD)may be obtained by cutting the fusing protein with the factor-Xa. Themethod have a high level expression and simple purification pmcess,which will be a verv usefulroad to eontribute and produce PTX-S3 in E. coli.
出处
《重庆医科大学学报》
CAS
CSCD
1996年第2期103-106,共4页
Journal of Chongqing Medical University
关键词
百日咳杆菌毒素
大肠杆菌
融合蛋白
基因
Bordetella peitussis toxin S3 subunit
Escherichia coli
Fusing protein