携带嗜热复制子的革兰氏阳性广宿主范围质粒的特性分析。

Characterization of gram-positive broad host-range plasmids carrying a thermophilic replicon.

作者信息

De Rossi E, Brigidi P, Rossi M, Matteuzzi D, Riccardi G

机构信息

Department of Genetics and Microbiology, A. Buzzati Traverso, University of Pavia, Pavia, Italy.

出版信息

Res Microbiol. 1991 May;142(4):389-96. doi: 10.1016/0923-2508(91)90108-m.

DOI:10.1016/0923-2508(91)90108-m

PMID:1908113

Abstract

The cryptic plasmid pBC1 (1.6 kb) isolated from Bacillus coagulans Zu1961 was genetically marked with the genes for chloramphenicol and ampicillin resistance (CmR and ApR) from the Escherichia coli plasmid pJH101. The recombinant vector obtained (pCP49, 7.0 kb) replicated and expressed CmR in B. subtilis and CmR and ApR in E. coli. Different shuttle vectors for Gram+ bacteria were also constructed by inserting pBC1 into the Staphylococcus aureus plasmid pC194. The smallest of these, pLM6 (2.8 kb), containing essentially pBC1 and the chloramphenicol acetyl transferase gene from pC194, replicated in B. subtilis at a copy number of 60. By electroporation, these plasmids were introduced and stably maintained in B. subtilis, B. amyloliquefaciens, S. aureus, S. carnosus and Lactobacillus reuteri.

摘要

从凝结芽孢杆菌Zu1961中分离出的隐蔽质粒pBC1（1.6 kb）用来自大肠杆菌质粒pJH101的氯霉素和氨苄青霉素抗性基因（CmR和ApR）进行了基因标记。获得的重组载体（pCP49，7.0 kb）在枯草芽孢杆菌中复制并表达CmR，在大肠杆菌中复制并表达CmR和ApR。通过将pBC1插入金黄色葡萄球菌质粒pC194中，还构建了用于革兰氏阳性菌的不同穿梭载体。其中最小的pLM6（2.8 kb），基本上包含pBC1和来自pC194的氯霉素乙酰转移酶基因，以60的拷贝数在枯草芽孢杆菌中复制。通过电穿孔，这些质粒被导入并稳定保存在枯草芽孢杆菌、解淀粉芽孢杆菌、金黄色葡萄球菌、肉葡萄球菌和罗伊氏乳杆菌中。