Detection Of White Spot Syndrome Virus (WSSV) And Expression Of Vp28 Gene

White spot syndrome virus is one of the main causes of shrimp diseases. We designed three pairs of primers in accordance with the sequence reported. We established an improved system of PCR for detecting WSSV in penaeid shrimp, that is a stability, rapidness, sensitivity and speciality system, including template preparation, the selection of primers and the improved parameters in PCR reaction.We also designed one pair of primes in accordance with the sequence reported for amplifying the total sequence of vp28 gene by PCR in the total DNA of virus isolated from Hainan and guangxi shrimp. The sequence analysis indicated that the two sequences seem similar and have no more difference. All kinds of isolates from different countries were compared through the software of DNAsisst and clustalxa 83.Results showed that the sequence of VP28 gene have little differences It is one highly conserved sequence in evolution .Furthermore,VP28 gene of WSSV was amplified by PCR according to the sequence reported,The VP28 gene is 615 nucleotides length and it was inserted into the prokaryotic expression vector pET-30a(+). Digestion identification and sequencing demonstrated that the recombinant vectors pET-30a-VP28 had been correctly constructed. The recombinant vectors were transformed into E.coli.BL21(DE3) respectively. Transformant were cultured and induced by addition of IPTG to a final concentration of 1mM. SDS-PAGE showed that theVP28 gene with pET-30a vector has been highly expressed in BL21(DE3).This study lay a firm foundation not only for preparation of antiserum later , subcellular vaccine of wssv, and detection wssv fast and effectively, but also for improving the methods to expression foreign gene.