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在渗透压稳定的培养基中用甘氨酸培养的乳球菌乳亚种经电穿孔高频转化。

High-Frequency Transformation, by Electroporation, of Lactococcus lactis subsp. cremoris Grown with Glycine in Osmotically Stabilized Media.

机构信息

Norwegian Dairies Association, Oslo, and Laboratory of Microbial Gene Technology, N-1432 As-NLH, Norway.

出版信息

Appl Environ Microbiol. 1989 Dec;55(12):3119-23. doi: 10.1128/aem.55.12.3119-3123.1989.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/64c7/203233/edfd53667ffe/aem00105-0098-a.jpg

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