工程菌种子制备方法对rEPA表达量影响的研究  

The effect of the recombinant bacteria seeds on the rEPA Expression quantity

作　　者：王永谦[1] 赵志强[1] 宣俊文[1] 谢贵林[1] 王燕[1] 杜琳[1] 

机构地区：[1]兰州生物制品研究所,730046

出　　处：《微生物学免疫学进展》2006年第2期12-14,共3页Progress In Microbiology and Immunology

摘　　要：由重组E.colirPE553D所表达的基因缺失突变脱毒的重组铜绿假单胞菌外毒素A(rEPA),目前大量以载体蛋白用于多种细菌多糖蛋白结合疫苗研究。rEPA的表达量受众多因素的影响,种子的制备方式就是重要因素之一。本文用长期冷冻保存的和新鲜提取的质粒分别转化宿主菌E.coliBL21(λDE3)后制备种子,并将它们和冻干保存的E.colirPE553D在相同条件下培养和诱导,经SDS-PAGE分析表明长期保存的质粒转化宿主后的重组工程菌生长速度较慢,但rEPA的表达量和新鲜质粒转化制备的工程菌基本相同,而冻干保存的工程菌E.colirPE553D几乎丧失了表达rEPA的能力。The recombinantpseudomonas aeruginosa exotoxin A（ rEPA）, expressed in E. coli BL21 （ λDE3 ） with a plasmid beating mutant EPA gene deleted the code for key amino acid 553D （glutamine 553 site） ,has been widely applied in polysaccharide-protein conjugate vaccine research. However, the expression of rEPA is affected by conditions which include the storage of plasmid and lyophilized recombinant bacteria. Plasmid stored at 4℃ over one year and freshly extract plasmid are transformed into E. coli BL21 （ λDE3 ） respectively. The two strains and lyophilized recombinant bacteria are cultured and induced in the same condition, then analysed by SDS-PAGE. Result shows： all freshly transformed strains have same expression quantity while the they have different growth rate, and the lyophilized strain almost lost the ability of expressing rEPA.

关 键 词：重组铜绿假单胞菌外毒素A 种子制备 表达量 工程菌 

分 类 号：R392[医药卫生—免疫学]