Characterization of degQ(Vh), a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain

doi:10.1128/AEM.00109-08

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	Characterization of degQ(Vh), a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain
	Zhang, Wei-wei 1,2; Sun, Kun 1,2; Cheng, Shuang 1,2; Sun, Li 1; Sun, L, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China
	2008-10-01
发表期刊	APPLIED AND ENVIRONMENTAL MICROBIOLOGY
ISSN	0099-2240
卷号	74 期号:20 页码:6254-6262
文章类型	Article
摘要	Vibrio harveyi is an important marine pathogen that can infect a number of aquaculture species. V. harveyi degQ (degQ(Vh)), the gene encoding a DegQ homologue, was cloned from T4, a pathogenic V. harveyi strain isolated from diseased fish. DegQ(Vh) was closely related to the HtrA family members identified in other Vibrio species and could complement the temperature-sensitive phenotype of an Escherichia coli strain defective in degP. Expression of degQVh in T4 was modulated by temperature, possibly through the sigma(E)-like factor. Enzymatic analyses demonstrated that the recombinant DegQVh protein expressed in and purified from E. coli was an active serine protease whose activity required the integrity of the catalytic site and the PDZ domains. The optimal temperature and pH of the recombinant DegQVh protein were 50 C and pH 8.0. A vaccination study indicated that the purified recombinant DegQVh was a protective immunogen that could confer protection upon fish against infection by V. harveyi. In order to improve the efficiency of DegQVh as a vaccine, a genetic construct in the form of the plasmid pAQ1 was built, in which the DNA encoding the processed DegQVh protein was fused with the DNA encoding the secretion region of AgaV, an extracellular beta-agarase. The E.coli strain harboring pAQ1 could express and secrete the chimeric DegQVh protein into the culture supernatant. Vaccination of fish with viable E. coli expressing chimeric degQ(Vh) significantly (P < 0.001) enhanced the survival of fish against V. harveyi challenge, which was possibly due to the relatively prolonged exposure of the immune system to the recombinant antigen produced constitutively, albeit at a gradually decreasing level, by the carrier strain.; Vibrio harveyi is an important marine pathogen that can infect a number of aquaculture species. V. harveyi degQ (degQ(Vh)), the gene encoding a DegQ homologue, was cloned from T4, a pathogenic V. harveyi strain isolated from diseased fish. DegQ(Vh) was closely related to the HtrA family members identified in other Vibrio species and could complement the temperature-sensitive phenotype of an Escherichia coli strain defective in degP. Expression of degQVh in T4 was modulated by temperature, possibly through the sigma(E)-like factor. Enzymatic analyses demonstrated that the recombinant DegQVh protein expressed in and purified from E. coli was an active serine protease whose activity required the integrity of the catalytic site and the PDZ domains. The optimal temperature and pH of the recombinant DegQVh protein were 50 C and pH 8.0. A vaccination study indicated that the purified recombinant DegQVh was a protective immunogen that could confer protection upon fish against infection by V. harveyi. In order to improve the efficiency of DegQVh as a vaccine, a genetic construct in the form of the plasmid pAQ1 was built, in which the DNA encoding the processed DegQVh protein was fused with the DNA encoding the secretion region of AgaV, an extracellular beta-agarase. The E.coli strain harboring pAQ1 could express and secrete the chimeric DegQVh protein into the culture supernatant. Vaccination of fish with viable E. coli expressing chimeric degQ(Vh) significantly (P < 0.001) enhanced the survival of fish against V. harveyi challenge, which was possibly due to the relatively prolonged exposure of the immune system to the recombinant antigen produced constitutively, albeit at a gradually decreasing level, by the carrier strain.
关键词	Heat-shock-protein Listeria-monocytogenes 10403s Cu/zn Superoxide-dismutase Escherichia-coli Survival Site-directed Mutagenesis Degp Protease Htra Family Elevated-temperatures Thermotoga-maritima Crystal-structure
学科领域	Biotechnology & Applied Microbiology ; Microbiology
DOI	10.1128/AEM.00109-08
URL	查看原文
收录类别	SCI
语种	英语
WOS记录号	WOS:000259985300011
引用统计	被引频次：123[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/1729
专题	实验海洋生物学重点实验室
通讯作者	Sun, L, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China
作者单位	1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
第一作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	Zhang, Wei-wei,Sun, Kun,Cheng, Shuang,et al. Characterization of degQ(Vh), a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain[J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY,2008,74(20):6254-6262.
APA	Zhang, Wei-wei,Sun, Kun,Cheng, Shuang,Sun, Li,&Sun, L, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China.(2008).Characterization of degQ(Vh), a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain.APPLIED AND ENVIRONMENTAL MICROBIOLOGY,74(20),6254-6262.
MLA	Zhang, Wei-wei,et al."Characterization of degQ(Vh), a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain".APPLIED AND ENVIRONMENTAL MICROBIOLOGY 74.20(2008):6254-6262.

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