Cloning And Expression Of Genes VP22/26 And VP28 Of White Spot Syndrome Virus

White spot disease(WSD)of penaeid shrimp is caused by white spot syndrome virus(WSSV),characterised by high and rapid mortality.WSSV has the widespread host range,including its sensitive host in the crustacean and in the insect outline animal,especially the decapoda class of shrimp and crab.Virions are rod-shaped to elliptical with a trilaminar envelope and they are large(80-120×250-380nm).WSSV is one kind of double-stranded DNA and its genome size is approximately 300kbp. WSSV is the main pathogen of wide area exploding shrimp virulent disease since 1993 in our country.WSSV propagates very quick,the infected shrimp would die in 3-7days and the mortality rate would reach as high as 90-100%,it would take the serious harm to our country and the world shrimp cultivation,but not yet has the effective prevention method at present.The envelope protein was the important detemining factor of virus infection and pathogenicity,WSSV envelope protein,the cell acceptor's relationship and the action mechanism were still not clear at present.WSSV has a main envelope protein VP28,envelope protein VP28 was proved that in virus's adsorption and the invasion process.At present t whether VP22/26 is the envelope or the nucleocapsid protein still has the dispute,virus's invasion needs viral and the cell surface acceptor's specificity union,but the envelope protein's specificity immune body and the viral granule' union has baffled the combine of viral and the cell acceptor's union,then has affected virus's adsorption,invasion or escapes the nucleocapsid process,some researches have already proofed that antiserum from thet rabbit immuned by WSSV envelope protein VP28 has the certainly neutral virus's ability in shrimp in vivo.Two pairs of primers were designed according to the sequences of genes VP22/26 and envelope genes VP28 of White spot syndrome virus(WSSV)in the GenBank.Using the primer 5.0 software to design and synthesizes two pairs of preimers.The two DNA fragments about 615bp and615bp amplified by PCR were linked to the BamH I restriction endonuclease site and cloned into E.colie expression vector pET-28a(+).Selects the recombinant and handwork withdraws the plasmid,after appraisal tramsformt the E.coli BL21.Carries on the sequence determination and the analysis.With IPTG induction at 30℃,the molecular weight of the engineered protein was about 28KDa,which was identified by SDS-PAGE analysis,were matched with anticipation.