谷氨酸棒杆菌的克隆载体系统
Cloning vector system for Corynebacterium glutamicum.
作者信息
Yoshihama M, Higashiro K, Rao E A, Akedo M, Shanabruch W G, Follettie M T, Walker G C, Sinskey A J
出版信息
J Bacteriol. 1985 May;162(2):591-7. doi: 10.1128/jb.162.2.591-597.1985.
Abstract
A protoplast transformation system has been developed for Corynebacterium glutamicum by using a C. glutamicum-Bacillus subtilis chimeric vector. The chimera was constructed by joining a 3.0-kilobase cryptic C. glutamicum plasmid and the B. subtilis plasmid pBD10. The neomycin resistance gene on the chimera, pHY416, was expressed in C. glutamicum, although the chloramphenicol resistance gene was not. The various parameters in the transformation protocol were analyzed separately and optimized. The resulting transformation system is simple and routinely yields 10(4) transformants per microgram of plasmid DNA.
摘要
通过使用谷氨酸棒杆菌-枯草芽孢杆菌嵌合载体，已开发出一种谷氨酸棒杆菌原生质体转化系统。该嵌合体通过连接一个3.0千碱基的谷氨酸棒杆菌隐蔽质粒和枯草芽孢杆菌质粒pBD10构建而成。嵌合体pHY416上的新霉素抗性基因在谷氨酸棒杆菌中表达，而氯霉素抗性基因则不表达。对转化方案中的各种参数进行了单独分析和优化。所得的转化系统简单，每微克质粒DNA通常可产生10⁴个转化体。
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