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IJFS 2014, 13 Back to browse issues page
Isolation and expression of recombinant viral protein (VP2) from Iranian isolates of Infectious Pancreatic Necrosis Virus (IPNV) in Escherichia coli
M Dadar *, H Rajabi Memari, R Peyghan, M.R Seifi Abad Shapouri
, dadar.m77@gmail.com
Abstract:   (4782 Views)

Infectious Pancreatic Necrosis Virus (IPNV) is a member of the family Birnaviridae that has been linked to high mortalities in salmonids. Bacterial based systems as live vectors for the delivery of heterologous antigens offer a number of advantages as vaccination strategies. VP2 is a structural viral protein of IPNV with immunogenicity effects. In this study IPNV was isolated from diseased fry of rainbow trout Oncorhynchus mykiss (Walbaum) using CHSE-214. Then an expression vector was constructed for expression of viral protein VP2. The designed vector was constructed based upon pET-26b (+) with T7 promoter. A fragment containing the full length of the VP2 gene of Iranian Sp strain was amplified by PCR using genomic RNA of IPNV as template and cloned inpET-26b(+) plasmid. Recombinant structural viral protein VP2 was expressed as a soluble, N-terminal PelB fusion protein and secreted into the periplasmic space of Escherichia coli BL21(DE3) and Rosetta (DE3). The glucose, Isopropyl-β-D- thiogalactopyranoside (IPTG) was used as a chemical inducer for rVP2 production in 37º C. The rVP2 was extracted from the periplasm by osmotic shock treatment. The presence of gene in bacterial system of E. coli was confirmed by gel electrophoresis technique. The constructed vector could efficiently express the rVP2 into the periplasmic space of E. coli. The successful cloning and expression of the structural viral protein gene into E. coli can be used for developing a useful and safe vaccine to control IPNV infection in Iranian fish industry.

Keywords: VP2, Infectious Pancreatic Necrosis Virus, Recombinant viral protein, Periplasmic space, Escherichia coli
Full-Text [PDF 380 kb]   (2035 Downloads)    
Type of Study: Orginal research papers | Subject: fish disease
Received: 2015/03/15 | Accepted: 2015/03/15 | Published: 2015/03/15
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Dadar M, Rajabi Memari H, Peyghan R, Seifi Abad Shapouri M. Isolation and expression of recombinant viral protein (VP2) from Iranian isolates of Infectious Pancreatic Necrosis Virus (IPNV) in Escherichia coli. IJFS. 2014; 13 (4) :856-868
URL: http://jifro.ir/article-1-1774-en.html

Volume 13, Issue 4 (2014) Back to browse issues page
مجله علوم شیلاتی ایران Iranian Journal of Fisheries Sciences
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