Construction and validation of a GFP-based vector for promoter expression analysis in the fish pathogen Flavobacterium psychrophilum
Construction and validation of a GFP-based vector for promoter expression analysis in the fish pathogen Flavobacterium psychrophilum
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DOI：
10.1016/j.gene.2012.01.069
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发表时间：
2012-04-15
期刊：
影响因子：
3.5
通讯作者：
Guijarro, Jose A.
中科院分区：
文献类型：
--
作者：
Gomez, Esther;Perez-Pascual, David;Guijarro, Jose A.
鱼类病原体嗜冷黄杆菌的研究由于难以对该生物体进行遗传操作而受到极大阻碍。尽管最近的出版物描述了通过转化和接合将遗传物质成功转移到这种细菌中，但仍然需要其他工具。本文报道了载体pCP 23-G的构建，它允许第一次监测转录调控在该病原体通过使用启动子gfpmut 3基因作为报告。此外，使用pCP 23-G能够对F. fpp 2-fpp 1、pdhB和gldj基因在不同生长条件下对嗜冷菌（psychrophilum）的影响。总体而言，pCP 23-G的构建有利于F.嗜冷菌，通过能够在体外和体内确定基因表达。此外，这也为研究这种细菌在鱼组织中的位置提供了可能性。(C)2012爱思唯尔有限公司版权所有。
The study of the fish pathogen Flavobacterium psychrophilum has been drastically hampered by the difficulty to perform genetic manipulation of this organism. Although recent publications described the successful transfer of genetic material into this bacterium by transformation and conjugation, additional tools are still needed. This paper reports the construction of vector pCP23-G, which permits for the first time to monitor transcriptional regulation in this pathogen by using a promoterless gfpmut3 gene as a reporter. Additionally, use of pCP23-G enabled the trancriptional analysis of three putative promoter regions of F. psychrophilum, corresponding to genes fpp2- fpp1, pdhB and gldj, under different growth conditions. Overall, the construction of pCP23-G facilitates genetic analysis in F. psychrophilum, by enabling the determination of gene expression both in vitro and in vivo. Furthermore, this would also open the possibility for studies on the location of this bacterium in the fish tissues. (C) 2012 Elsevier B.V. All rights reserved.