Norwegian Dairies Association, Oslo, and Laboratory of Microbial Gene Technology, N-1432 As-NLH, Norway.
Appl Environ Microbiol. 1989 Dec;55(12):3119-23. doi: 10.1128/aem.55.12.3119-3123.1989.
An efficient method for genetic transformation of lactococci by electroporation is presented. Highly competent lactococci for electrotransformation were obtained by growing cells in media containing high concentrations of glycine and 0.5 M sucrose as the osmotic stabilizers. These cells could be stored at -85 degrees C without loss of competence. With Lactococcus lactis subsp. cremoris BC101, a transformation frequency of 5.7 x 10 transformants per mug of pIL253 DNA was obtained, which represents 5% of the surviving cells. All the lactococcal strains tested could be transformed by the present method.
本文介绍了通过电穿孔高效转化乳球菌的方法。通过在含有高浓度甘氨酸和 0.5 M 蔗糖的培养基中培养细胞,获得了高电转化能力的乳球菌作为渗透稳定剂。这些细胞可以在-85°C下储存而不会丧失转化能力。对于 Lactococcus lactis subsp. cremoris BC101,每微克 pIL253 DNA 可获得 5.7 x 10 个转化体,占存活细胞的 5%。本方法可用于转化所有测试的乳球菌菌株。