乳杆菌电转化条件的研究  

The Factors Affected Transformation Efficiency of Lactobacillus by Electroporation

作　　者：贾士芳[1] 王荫榆[1] 郭兴华[1] 还连栋[1] 

机构地区：[1]中国科学院微生物研究所

出　　处：《生物工程学报》1998年第4期429-433,共5页Chinese Journal of Biotechnology

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８７５ｋＶ／ｃｍ，电阻１００Ω或２００Ω，电容２５μＦ和２×磷酸缓冲液。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.

关 键 词：乳杆菌 质粒 电转化 

分 类 号：Q78[生物学—分子生物学] Q939.117