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Epidemiological Investigation of TS and IHHN and Preliminary Study on Interaction of Structural Proteins of TSV VP1 and VP3

Author: LiMing
Tutor: FeiRongMei
School: Nanjing Agricultural College
Course: Preventive Veterinary Medicine
Keywords: IHHN TSV Epidemiological investigation VP1 VP3 Proteins interaction
CLC: S945
Type: Master's thesis
Year: 2011
Downloads: 1
Quote: 0
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Abstract


Taura Syndrome is a serious viral disease harmful shrimp farming.It is caused by taura syndrome virus(TSV). Penaeus vannamei and P. stylirostris are the main host of tsv,which caused high burst rate and death rate.Infectious hypodermal and hematopoietic necrosis virus(IHHNV) is one of the highly pathogenic shrimp viruses causing disease in shrimp.The host range of IHHNV is very wide,the main host of IHHNV is Penaeus stylirostris. In this paper, we investigate TSV and IHHNV of shrimp in region of Jiangsu Province.In order to investigate TSV and IHHNV of shrimp in region of Jiangsu Province 718 shrimp samples of shrimp were collected from different areas of Jiangsu Province,such as Yancheng,Lianyungang and Nanjing.The shrimp samples were detected for TSV and TSV infection by LAMP and PCR.The results showed that there were zero TSV and a high prevalence of IHHNV(20.0%).TSV contains three major polypeptides VP1,VP2 and VP3(55,40and 24kD) and one minor polypeptides VP0(58kD) constitude its proteinic capsid.In this paper, recombinant protein VP1 and VP3 were expressed to research preparation of Taura VP1 and VP3 Virus-like particles.Based on the genome sequence of Chinese variant ZHZC3,primer were designed to amplify VP1 and VP3 gene.Target genes were got by RT-PCR. The products were inserted to plasmid pColdTMTF for making recombinant plasmid pCold-VP1、pCold-VP3, then transformed to host strain BL21(DE3). The positive clone identified by endonuclease digest were induced by IPTG. Detection target proteins by SDS-PAGE. Both of recombinant are well expressed in the form of supernatant with 1.0 mmol/L IPTG at 15℃. An Ni affinity column is used to purify the inclusion body protein. After negative staining by transmission electron microscopy, VP1 and VP3 proteins were not self-assemble into virus-like particles.Based on the genome sequence of Chinese variant ZHZC3,primer were designed to amplify VP1 gene by RT-PCR. The product was inserted to plasmid pGEX-4T-3 for making recombinant plasmid and then transformed to host strain BL21. The positive clone identified by endonuclease digest were induced by IPTG..Pull-Down assay analyze the interaction between pCold-VP3 and pGEX-VP1 recombinant protein, the results showed that they did not have interactions.

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Fisheries Protection > Pest and Disease Control of the crustaceans
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