摘要
构建了荧光假单胞菌工程菌的转化系统,探索了多寄主质粒pSUP 106在不同CaCl_2浓度、不同热激时间和荧光假单胞菌受体在不同生长期的转化频率,建立了一个简便可行的荧光假单胞菌转化方法.结果表明,在荧光假单胞菌Pfx-18生长到0 D_(600)≈0.55时,用25mmol/L CaCl_2处理细胞,热激2min,转化频率最高,可达10~5/ng DNA.同时讨论了Mn^(2+)和Mg^(2+)对转化频率的影响,以及电转化的效果,为进一步利用该转化系统进行异源基因的克隆与表达研究奠定了基础.
In order to establish a transformation system for Pseudomonas fluorescens,different factors which effect transformation frequency of Pseudomonas fluorescenswere tested. The result showed that transformation frequency could acheive 105 cells/μg DNA when OD600 of Pseudomonas fluorescens was 0.55 and the cells were treated with 25mmol/L CaCl2 and heated for 2 minuts at 42℃. The effect of Mg2+and Mn2+ on transformation frequency were also discussed. The system could beuseful for the study of cloning and expression of heterlogous genes in P. fluorescens.
出处
《微生物学报》
CAS
CSCD
北大核心
1995年第4期250-253,共4页
Acta Microbiologica Sinica
基金
国家自然科学基金资助项目
