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。
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.
出处
《生物工程学报》
CAS
CSCD
北大核心
1998年第4期429-433,共5页
Chinese Journal of Biotechnology
关键词
乳杆菌
质粒
电转化
Lactobacillus ,plasmid,electrotransformation
