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口腔链球菌NCTC 11427基因转化的特性:使用新型穿梭载体在口腔链球菌中表达肺炎球菌酰胺酶

Characterization of genetic transformation in Streptococcus oralis NCTC 11427: expression of the pneumococcal amidase in S. oralis using a new shuttle vector.

作者信息

Ronda C, García J L, López R

机构信息

Centro de Investigaciones Biológicas C.S.I.C., Madrid, Spain.

出版信息

Mol Gen Genet. 1988 Dec;215(1):53-7. doi: 10.1007/BF00331302.

Abstract

We have worked out conditions for the study of competence development and genetic transformation in Streptococcus oralis NCTC 11427 (type strain), a species that contains choline in the cell wall. The peak of competence was found at the early exponential phase of growth and the optimal conditions for transformation were achieved with shuttle plasmids prepared from S. pneumoniae or from Escherichia coli serving as donor DNA. Transformation with dye-buoyant density gradient purified plasmid preparations followed first-order kinetics. The pneumococcal amidase can be expressed in S. oralis harbouring a plasmid carrying the lytA gene. This enzyme lysed the cell wall of the transformed cell in the presence of detergents.

摘要

我们已经确定了用于研究口腔链球菌NCTC 11427(模式菌株)感受态发育和遗传转化的条件,该菌种的细胞壁中含有胆碱。发现感受态高峰出现在生长的指数早期阶段,使用从肺炎链球菌或大肠杆菌制备的穿梭质粒作为供体DNA可实现最佳转化条件。用染料浮力密度梯度纯化的质粒制剂进行转化遵循一级动力学。肺炎球菌酰胺酶可在携带lytA基因的质粒的口腔链球菌中表达。这种酶在去污剂存在下可裂解转化细胞的细胞壁。

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