重组人内皮抑素工程菌菌种稳定性考察  

Stability of recombinant human endostatin engineering bacteria E.Coli

作　　者：陈罗胜[1] 殷润婷[1] 徐寒梅[1] 奚涛[1] 

机构地区：[1]中国药科大学生物技术中心,南京210009

出　　处：《生物加工过程》2008年第6期58-62,共5页Chinese Journal of Bioprocess Engineering

摘　　要：研究重组人内皮抑素突变体质粒HM-E的宿主菌E.coliBL21(DE3)在LB培养基中传代50代过程中菌种的稳定性。研究表明,重组人内皮抑素工程菌在传代50代的过程中质粒稳定性(ST)高,为98%,菌体与菌落呈典型的大肠杆菌形态,反复冻融后表达量未下降,目标蛋白在菌体超声的沉淀中质量分数为(58.75±3.78)%,同时,四甲基偶氮唑盐微量反应比色法(MTT)结果表明目的蛋白质量浓度为20μg/mL时对内皮细胞ECV-304具有很高的抑制率,达(94.72±2.10)%。The strain stability of recombinant E. coli BL21 （DE3） cultured in LB was studied by bacterial morphology, plasmid stability （ST）, restriction enzyme digestion analysis of plasmid, expression of rhEndostatin, and expression activity after freeze thawing. The plasmid of the bacteria was kept well after 50 generations and ST of 50th generation remained 98%. In the 50th generation, the cell and the colonial morphology were identical with that of the typical E. coli strain. The mass fraction was （ 58.75 ± 3.78 ） % after freeze thawing. MTT demonstrated that the target protein of 20 μg/mL had high inhibition on ECV-304 and reached （94. 72 ± 2. 10）%, thus recombinant E. coli BL21 （DE3） was stable.

关 键 词：工程菌 质粒稳定性 重组人内皮抑素 菌种传代 

分 类 号：Q78[生物学—分子生物学]