重组水蛭素Ⅲ工程菌菌种稳定性考察  

Study of Stability in Recombinant Hirudin Ⅲ-Excreting Engineering Bacteria

作　　者：韦利军[1] 阮期平[1] 郭海[2] 吴梧桐[1] 

机构地区：[1]中国药科大学生物制药学院,江苏南京210009 [2]北京林业大学生物科学与技术学院,北京100083

出　　处：《药物生物技术》2004年第2期86-90,共5页Pharmaceutical Biotechnology

摘　　要：研究基因工程菌E .coliAS1.35 7在LB培养基中传代 5 0代过程中菌种的稳定性。以菌体和菌落的形态、质粒稳定性 (ST)、质粒酶切图谱和水蛭素的表达量以及比活等方面进行考察 ,以评价工程菌的稳定性。重组水蛭素Ⅲ工程菌在传代 5 0代的过程中质粒稳定性高 ,传代 10代内质粒稳定性 (ST)为 98% ,且表达稳定 ,表达产物可达发酵液可溶性蛋白总量的 6 0 % ,产物的抗凝血酶活力大于 2 .0× 10 3 ATU/ml,传代 5 0代的菌体与菌落呈典型的大肠杆菌形态。重组水蛭素Ⅲ工程菌菌种稳定。Strain stability of recombinant E.coli AS1.357 when cultured in LB substrate was studied. The stability was evaluated by studying the bacterial morphology, plasmid stability (ST), restriction enzyme digestion analysis of recombinant plasmids, expression of hirudin Ⅲ and anti-thrombin activity. The plasmid of this bacteria was kept well in 50 generations and its stability was high, in 10 generations the ST was higher than 98%. The expression was stable and the anti-thrombin activity kept to be stable, of about 2.0×10 3 ATU/ml. In 50 generations the bacteria's state of body and colony were all typical. Recombinant bacteria E.coli AS1.357 is stable and is suitable for industrialized production.

关 键 词：重组水蛭素 质粒稳定性 工程菌稳定性 

分 类 号：Q78[生物学—分子生物学]