摘要
用SDS处理谷氨酸棒杆菌1014(Corynebactertum glutamlcum 1014),获得消除质粒的衍生株1014-6。通过质粒pXZ10145(Cm^r)转化1014-6菌株的原生质体,研究了转化最适条件。0.6u/ml青霉素处理对数期菌体可明显提高转化效率。转化促进剂PEG以分子量6000、浓度30%为最佳。转化在37℃水浴进行3min效果最好。转化效率最高可达2×10~4转化子/μg DNA。质粒pXZ10145也已成功地转入钝齿棒杆菌B9(C.crenatum B9)。并在新宿主中基本保持稳定。
A transformation system in Corynebacterium was developed C.glutamicum 1014-6 containing no plasmid was derived from C. glutamicum 1014 which harbors two kind of plasmids after the treatment with SDS on C.glutamicum 1014. Optimal conditions of protoplast transformation of C.glutamicum 1014-6 with plasmid pXZ10145(C_m^r) were studied. Protoplast transformation efficiency is distinctly increased by the treatment of early logarithmic phase bacterial cells with penicillin G.Optimal transformation conditions are 30% PEG 6000 and 37℃ water bath for 3 minutes, yielding 2×10~4 transformants per μg of DNA.Plasmid pXZ10145 originally from C.glutamicum 1014, could transform C.crenatum B9 with an efficiency of 3.3×10~3 transformants per μg of DNA. The plasmid was stably maintained in the new host.
出处
《生物工程学报》
CAS
CSCD
北大核心
1989年第1期51-56,共6页
Chinese Journal of Biotechnology
关键词
棒杆菌
原生质体
质粒转化
Corynebacterium
protoplast
plasmid pXZ10145
transformation
