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乳杆菌电转化条件的研究
The Factors Affected Transformation Efficiency of Lactobacillus by Electroporation
【摘要】 用pGK12、pMG36C等质粒电转化不同的乳杆菌。研究了影响转化效率的多种因素。受体细胞经20μg/ml氨苄青霉素处理1h,可以提高转化效率200倍。同时,发现电击后的细胞必需在高渗培养基中才能存活,电击后2~3h的复苏表达期和用亚抑制抗生素浓度选择转化子,这些对电转化成功以及提高转化效率都是十分关键的。在改进电转化方法中,各种参数为电场强度875kV/cm,电阻100Ω或200Ω,电容25μF和2×磷酸缓冲液。
【Abstract】 Plasmid pGK12,pHPSOD DNA were transformed into different kinds of Lactobacillus strains by electroporation.A number of factors that affected transformation efficiency were investigated.Treatment of the recipient Lactobacilli with ampicillin improved electrotransformation efficiencies up to 200 fold.A post pulse recovery time of 2~3h and the use of sub inhibitory concentrations of antibiotics in the selective plates were very critical for electrotransformation.Several parameters in improved protocol were voltage 8,75kV.cm -1 ,resistance 100Ω,capacitance 25μF,buffer 2×PEB1.
- 【文献出处】 生物工程学报 ,CHINESE JOURNAL OF BIOTECHNOLOGY , 编辑部邮箱 ,1998年04期
- 【分类号】Q789,
- 【被引频次】77
- 【下载频次】632