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用于嗜酸乳杆菌启动子分析的mCherry蛋白载体的构建与验证

Construction and validation of a mCherry protein vector for promoter analysis in Lactobacillus acidophilus.

作者信息

Mohedano M Luz, García-Cayuela Tomás, Pérez-Ramos Adrián, Gaiser Rogier A, Requena Teresa, López Paloma

机构信息

Departamento de Microbiología Molecular y Biología de las Infecciones, Centro de Investigaciones Biológicas (CIB-CSIC), Ramiro de Maeztu 9, 28040, Madrid, Spain.

出版信息

J Ind Microbiol Biotechnol. 2015 Feb;42(2):247-53. doi: 10.1007/s10295-014-1567-4. Epub 2014 Dec 23.

Abstract

Lactobacilli are widespread in natural environments and are increasingly being investigated as potential health modulators. In this study, we have adapted the broad-host-range vector pNZ8048 to express the mCherry protein (pRCR) to expand the usage of the mCherry protein for analysis of gene expression in Lactobacillus. This vector is also able to replicate in Streptococcus pneumoniae and Escherichia coli. The usage of pRCR as a promoter probe was validated in Lactobacillus acidophilus by characterizing the regulation of lactacin B expression. The results show that the regulation is exerted at the transcriptional level, with lbaB gene expression being specifically induced by co-culture of the L. acidophilus bacteriocin producer and the S. thermophilus STY-31 inducer bacterium.

摘要

乳酸杆菌在自然环境中广泛存在,并且作为潜在的健康调节因子越来越受到研究。在本研究中,我们对广宿主范围载体pNZ8048进行了改造,使其表达mCherry蛋白(pRCR),以扩大mCherry蛋白在乳酸杆菌基因表达分析中的应用。该载体也能够在肺炎链球菌和大肠杆菌中复制。通过表征乳酸杆菌素B表达的调控,验证了pRCR作为启动子探针在嗜酸乳杆菌中的应用。结果表明,这种调控发生在转录水平,嗜酸乳杆菌细菌素产生菌与嗜热链球菌STY-31诱导菌共培养可特异性诱导lbaB基因表达。

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