摘要
目的观察甲基化缺陷的大肠杆菌DM1菌株与普通DH5α菌株感受态转化效率的差别。方法用常规氯化钙法制备二者的感受态细胞,质粒转化后计算转化率。结果DM1菌株感受态细胞转化率为(2.2±0.3)×106CFU/μg,DH5α菌株感受态细胞转化率为(6.3±0.5)×106CFU/μg。结论成功建立一种高效制备甲基化缺陷菌株DM1感受态细胞的方法,虽然该法制备DM1感受态细胞转化效率比DH5α低2-3倍,但其能够高效满足质粒克隆实验中Dam或Dcm甲基化的限制性位点的使用要求。
Objective To compare the difference of transformation between demethylation E.coli DM1 and DH5α competent cells.Methods Demethylation E.coli DM1 and DH5α competent cells were prepared by the routine CaCl2 method,and then the transformation of competent cells was calculated.Results The transformation was（2.2±0.3）×106 CFU/μg,（6.3±0.5）×106 CFU/μg in demethylation E.coli DM1 and DH5α competent cells,respectively.Conclusion An efficient and rapid method for the preparation of demethylation E.coli DM1 competent cells is successfully established,which can be efficiently utilized in molecular cloning for Dam or Dcm restriction sites.
出处
《山西医科大学学报》
CAS
2010年第4期322-324,共3页
Journal of Shanxi Medical University
基金
山西省实验动物科研专项基金资助项目[2008k02]
关键词
感受态细胞
甲基化
DM1
DH5Α
转化率
competent cell
methylation
E.coli DM1
E.coli DH5α
transformation
