摘要
目的优化高转化效率甲基化酶缺陷的大肠杆菌DM1感受态细胞的制备条件。方法采用不同生长状态、转化冰浴时间、热激时间及转化后复苏时间的大肠杆菌DM1制备感受态细胞,分析转化效率。结果 A600值为0.39和0.69的菌液制备感受态细胞,转化冰浴时间为30min,42℃热激处理90s,大肠杆菌DM1的转化效率最高;转化后复苏时间对细菌转化效率的影响不明显。结论已优化了高转化效率甲基化酶缺陷的大肠杆菌DM1感受态细胞的制备条件,可满足大部分在质粒中进行的常规克隆的需要。
Objective To optimize the condition for preparation of competent cells of methylase-deficiency E.coli DM1 with high transformation efficiency.Methods The effect of growth state of E.coli DM1,time for ice bath,time for heat shock and resuscitation time after transformation on transformation efficiency were observed,based on which the condition for preparation of competent cells was optimized.Results The transformation efficiency of competent cells prepared with E.coli DM1 at A600 of 0.39 and 0.69 kept in ice bath for 30 min and incubated at 42℃ for 90 s for heat shock was satisfactory.However,the resuscitation time after transformation showed no significant effect on transformation efficiency.Conclusion The condition for preparation of competent cells of methylase-deficiency E.coli DM1 with high transformation efficiency was optimized,which met most of the requirements for routine cloning in plasmids.
出处
《中国生物制品学杂志》
CAS
CSCD
2010年第5期533-535,538,共4页
Chinese Journal of Biologicals
关键词
大肠杆菌
感受态细胞
转化效率
E. coli
Competent cells
Transformation efficiency
