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Functional Analysis of Envelope Protein VP33 and Identification of Non-structure Protein Wsv406 of White Spot Syndrome Virus

Author: LinYi
Tutor: YangFeng
School: Xiamen University
Course: Biochemistry and Molecular Biology
Keywords: WSSV VP33 ORF wsv406
CLC: S945
Type: Master's thesis
Year: 2008
Downloads: 43
Quote: 0
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Abstract


WSSV (white spot syndrome virus, WSSV) is one of the major viral pathogens that endanger the world's shrimp farming industry . It is a capsule , inclusion bodies , the rod - like double-stranded circular DNA virus , with infection and strong high mortality , difficult to control . Due to its wide host range , infecting a variety of crustaceans , are posing a serious threat to aquaculture and the marine environment . This thesis is focused on the research of virus structural proteins . This is conducive to reveal the assembly mechanism in the viral replication process , so as to provide a strong scientific basis for the eventual establishment of effective prevention and treatment methods . The major research work is divided into two parts: (1) the identification of the membrane protein VP33 . Adopted by the previous studies on the structure of the whole virus proteins by SDS-PAGE and mass spectrometry , the combination of amino acid sequence analysis , has been identified VP33 WSSV genome ORF wsv254 encoded membrane protein . In E.coli XL 1 - Blue and E : coliBL21 ( DE3 ) expression and purification of recombinant VP33 protein with a 6 × His tag and GST expression , and to collect the purified fusion protein (( His) 6 -VP33) renaturation after injection the mice that received specific polyclonal antibody . Thereafter, the use of GST pull-down analysis and Far-Western blot analysis proved the presence of a specific interaction between the VP33 with itself and another major membrane protein VP24 . (2) analysis of by WSSV whole genome sequencing , we obtained a the ORF wsv406 DNA sequence . This article cloned wsv406 whole sequence of genes to the vector pGEX - 2TH and expressed in E.coli BL21 (DE3) , the form of inclusion body insoluble fusion protein . Inclusion body refolding Reactive Protein purification collected after the injection mice were specific polyclonal antibody .

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