Analysis,cloning,expression of CGSIV ORF22R in E.coli
Author(s): ZHANG Xin, ZHAO Gaoxiang, LI Wei, ZHOU Tianhong
Pages: 118-123
Year: 2014 Issue: 2
Journal: Journal of Jinan University（Natural Science & Medicine Edition）
Keyword: Chinese giant salamander iridovirus; sequence analysis; prokaryotic ex-pression;
Abstract: Recently,Infectious diseases are implicated in the declines and the economic loss in cul-tured and wild Chinese giant salamander.One of pathogens has been recognized as Chinese giant sala-mander iridovirus (CGSIV).The putative ORF22R of CGSIV was amplified and sequenced.The ORF consists of 1 818 bp,which codes for a protein of 605 aa with a predicted molecular mass of 65.185 kD. Comparative studies of the identity of the amino acid sequence of ORF22R were carried out with different iridovirus species.Alignment of the identity of the amino acid sequence of this ORF of CGSIV and amphib-ian-like ranaviruses(ALRV)showed 95.9%-98.2% identity,but significantly higher than that of GIV-like viruses (31.7%)and the Lymphocystis disease virus 1 and C in other genus of Ranavirus (~16%). ORF22R was cloned into the pET22b vector,and expressed in E.coli Rosetta.The protein ORF22R was purified with nickel column chromatography after inclusion body lysis,and isolated by SDS-PAGE.
Pages: 118-123
Year: 2014 Issue: 2
Journal: Journal of Jinan University（Natural Science & Medicine Edition）
Keyword: Chinese giant salamander iridovirus; sequence analysis; prokaryotic ex-pression;
Abstract: Recently,Infectious diseases are implicated in the declines and the economic loss in cul-tured and wild Chinese giant salamander.One of pathogens has been recognized as Chinese giant sala-mander iridovirus (CGSIV).The putative ORF22R of CGSIV was amplified and sequenced.The ORF consists of 1 818 bp,which codes for a protein of 605 aa with a predicted molecular mass of 65.185 kD. Comparative studies of the identity of the amino acid sequence of ORF22R were carried out with different iridovirus species.Alignment of the identity of the amino acid sequence of this ORF of CGSIV and amphib-ian-like ranaviruses(ALRV)showed 95.9%-98.2% identity,but significantly higher than that of GIV-like viruses (31.7%)and the Lymphocystis disease virus 1 and C in other genus of Ranavirus (~16%). ORF22R was cloned into the pET22b vector,and expressed in E.coli Rosetta.The protein ORF22R was purified with nickel column chromatography after inclusion body lysis,and isolated by SDS-PAGE.
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