title: Development of a regulatable expression system for the functional study of Vibrio vulnificus essential genes.
abstract: We developed a regulatable gene expression system for Vibrio vulnificus, which contains a lacIq-pTrc cassette. Monomeric red fluorescence protein (mRFP) was used as a reporter to test this system. The results showed that this system tightly controlled the expression of mRFP without leaky expression and was suitable for the controlled expression of genes encoding recombinant proteins in V. vulnificus. To demonstrate the utility of this system, a dominant negative form of V. vulnificus VVMO6_RS04990, a homolog of Escherichia coli LolD that is essential in lipoprotein transport and membrane biogenesis, was inducibly expressed. Expression of the dominant negative LolD homolog, which has a mutation in the ATPase domain, resulted in a growth defect in V. vulnificus cells and impaired cell envelope stability. This result suggests that the V. vulnificus LolD homolog plays a role in cell envelope biogenesis. This tight and titratable expression system will therefore be a valuable tool for the study of essential genes in V. vulnificus.
authors.affiliation: Shandong Province Key Laboratory of Applied Mycology, School of Life Sciences, Qingdao Agricultural University, Qingdao, 266109, Shandong, People's Republic of China.
authors.forename: Yan-Jiao
authors.initials: YJ
authors.lastname: Zhang
authors.affiliation: Shandong Province Key Laboratory of Applied Mycology, School of Life Sciences, Qingdao Agricultural University, Qingdao, 266109, Shandong, People's Republic of China.
authors.forename: Guozhong
authors.initials: G
authors.lastname: Chen
authors.affiliation: Shandong Province Key Laboratory of Applied Mycology, School of Life Sciences, Qingdao Agricultural University, Qingdao, 266109, Shandong, People's Republic of China.
authors.forename: Huiyuan
authors.initials: H
authors.lastname: Lin
authors.affiliation: Shandong Province Key Laboratory of Applied Mycology, School of Life Sciences, Qingdao Agricultural University, Qingdao, 266109, Shandong, People's Republic of China.
authors.forename: Pan
authors.initials: P
authors.lastname: Wang
authors.affiliation: Shandong Province Key Laboratory of Applied Mycology, School of Life Sciences, Qingdao Agricultural University, Qingdao, 266109, Shandong, People's Republic of China.
authors.forename: Baozhi
authors.initials: B
authors.lastname: Kuang
authors.affiliation: Central Laboratory, Qingdao Agriculture University, Qingdao, 266109, Shandong, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: Shandong Province Key Laboratory of Applied Mycology, School of Life Sciences, Qingdao Agricultural University, Qingdao, 266109, Shandong, People's Republic of China. shiyongchen@hotmail.com.|Collaborative Innovation Center for Marine Biomass Fibers, Materials and Textiles of Shandong Province, Qingdao University, Qingdao, 266071, Shandong, People's Republic of China. shiyongchen@hotmail.com.
authors.forename: Shiyong
authors.identifier: http://orcid.org/0000-0002-2616-4367
authors.initials: S
authors.lastname: Chen
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D041261:Vibrio vulnificus / Q000235:genetics*; D000097573:Red Fluorescent Protein
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C407983:LolD protein, E coli
keywords: Essential gene; Expression system; Functional study; Regulatable; Vibrio vulnificus