RESEARCH ON THE PROKARYOTIC EXPRESSION AND CHARACTERISTICS OF VP28,A STRUCTURE PROTEIN OF PENAEUS MONODON WHITE SPOT SYNDROME VIRUS
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Abstract
White Spot Syndrome Virus(WSSV)is an enveloped DNA virus,which causes 100% cumulative mortality of cultured shrimps in 3 4 days. The virus particles contain five major proteins:VP28、VP26、VP24、VP19 and VP15. The mode of entry and systemic infection of WSSV in shrimp and the functions of these virion proteins are not clear. But more and more attention is paid to the function of VP28,which is likely to play a key role in the initial steps of WSSV infection in shrimps. Towards simplifying to study the role of VP28 in the interaction between WSSV and host cells,the gene of VP28 has been cloned into a prokaryotic expression system,induced and expressed for further researching its characteristics.This prokaryoti expression product could not secrete out of the cell membrane,because it is a membrane protein. Rabbit antiserum against prokaryotic expressed VP28 could recognize both prokaryotic expressed VP28 and native WSSV;Mouse antiserum against native WSSV could recognize both WSSV and prokaryotic expressed VP28. A scFv antibody, designated B7. selected from phage display antibody library against WSSV by panning with prokaryotic expressed VP28,could also recognize both prokaryotic expressed VP28 and native WSSV. These data show that the VP28 produced in a prokaryotic expression system has analogous immunogenic characteristics to its native form,although their protein post modification may differ.
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