Study On Basic Properties Of White Spot Syndrome Virus And Function Of Protein Kinase Wsv083

Shrimp white spot syndrome virus (white spot syndrome virus, WSSV) is arod-shaped DNA virus, with the envelop and the tegument, DNA as double chainring. WSSV is currently the main diseases of prawn aquaculture, caused massiveharm. Therefore, the study on properties and infection mechanism of WSSV is ofgreat significance for prawn aquaculture.The main content of this study are as follows:①determination of WSSV particle mass and its basic components;②investigate the role of the microbial infection in the death of crayfishinfected with WSSV;③the identification of the WSSV particle surface protein;④a WSSV protein kinase wsv083were studied.The results are as follows:First the mass of WSSV genome was calculated, and the proportion of thegenomic DNA accounted for total WSSV particle virus mass was also measured,finally calculate the mass of a complete WSSV particle. The relative proportionsof the major envelop protein were also studied. WSSV suspension for each of theOD600was equivalent to3.34×108/L of the virus concentration.In addition,WSSV components in total weight in proportion through the determination wereas follows: the genomic DNA accounted for10.17%, nucleocapsid protein is43.07%, envelop lipid content is22%, envelop protein content is24.76%.To study the role of microorganism in the process of death of theProcambarus clarkia infected with WSSV(white spot syndrome virus), themortality rate of Procambarus clarkia injected with antibiotics decreasedsignificantly compared with the control group. Plate count showed that thehemolymph of Procambarus clarkia infected with WSSV have significantly increased the number of microorganisms. Analysis showed that they wereAeromonas hydrophila, Citrobacter freundii, Stenotrophomonas maltophilia andAcinetobacter junii. All of them were opportunistic pathogen. Antibiotic had nosignificant effect on the WSSV’s proliferation.The results showed that thedisappearance of the blood cells of Procambarus clarkia infected with WSSV invivo led to a substantial increase in the number of pathogenic microorganisms,thereby accelerating the rate of Procambarus clarkia deaths.Using the biotin label transfer technique, we found that the biotin label wastransferred from BSA or gill member protein of Litopenaeus vannamei to theenvelop protein of WSSV. Western blotting showed that two proteins whosemolecular weight among50-55KD were labeled with biotin. It was concludedthat the two proteins was location on the surface of the WSSV viron and couldinteract with the gill member protein of Litopenaeus vannamei. They wereestimated to be VP52A and VP56.VP28、VP19and wsv083were expressed in E. coli and purified. They wereput in kinase buffer to learn if wsv083can catalyze VP28and VP19’sphosphorylation. Results showed that wsv083cann’t. We also found that therewere threonine and tyrosine phosphorylation but no serine phosphorylation inwsv083. And wsv083mutant was not phosphorylated. It was concluded thatwsv083catalyze autophosphorylation. But its specific function needs furtherstudy.