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通过电穿孔法用质粒pLHR对瑞士乳杆菌嗜酸奶亚种进行转化。

Transformation of Lactobacillus helveticus subsp. jugurti with plasmid pLHR by electroporation.

作者信息

Hashiba H, Takiguchi R, Ishii S, Aoyama K

机构信息

Technical Research Institute, Snow Brand Milk Products Co., Ltd., Saitama, Japan.

出版信息

Agric Biol Chem. 1990 Jun;54(6):1537-41.

PMID:1368569
Abstract

Lactobacillus helveticus subsp. jugurti was transformed with plasmid, pLHR (8.5 kilobases), by electroporation. The plasmid, pLHR, consists of a cryptic plasmid, pLJ1, from L. helveticus subsp. jugurti, the Escherichia coli vector pBR329, and the erythromycin resistance gene of pAM beta 1 from Enterococcus faecalis. Maximum transformation efficiency of 1.3 x 10(4) transformants per microgram of DNA was obtained by exposure to a pulse with an exponential decay waveform at 4 kV/cm with 25 microF capacitance. The presence of glycine in the growth medium was essential for transformation. Plasmid DNA isolated from transformants had not undergone detectable rearrangements or deletions. In addition, it was found that L. helveticus subsp. jugurti has a restriction and modification system.

摘要

瑞士乳杆菌嗜酸奶亚种通过电穿孔法用质粒pLHR(8.5千碱基)进行转化。质粒pLHR由来自瑞士乳杆菌嗜酸奶亚种的隐蔽质粒pLJ1、大肠杆菌载体pBR329以及来自粪肠球菌的pAMβ1的红霉素抗性基因组成。通过在4 kV/cm、25 μF电容下用指数衰减波形的脉冲进行处理,获得了每微克DNA 1.3×10⁴个转化体的最大转化效率。生长培养基中甘氨酸的存在对转化至关重要。从转化体中分离出的质粒DNA未发生可检测到的重排或缺失。此外,还发现瑞士乳杆菌嗜酸奶亚种具有限制和修饰系统。

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