PROCESSING OF THE DENGUE VIRUS TYPE-2 PROTEINS PRM AND C-PRM

被引:70
作者
MURRAY, JM
AASKOV, JG
WRIGHT, PJ
机构
[1] MONASH UNIV,DEPT MICROBIOL,CLAYTON,VIC 3168,AUSTRALIA
[2] QUEENSLAND UNIV TECHNOL,CTR MOLEC BIOTECHNOL,BRISBANE,QLD 4001,AUSTRALIA
关键词
D O I
10.1099/0022-1317-74-2-175
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A glycoprotein C-prM of 35000 M(r) was immunoprecipitated from lysates of Aedes albopictus cells infected with dengue virus type 2 (DEN-2) using antisera directed against the C protein or an amino-terminal fragment of the prM glycoprotein. C-prM was not detected in infected Vero cells. The prM glycoprotein synthesized in infected A. albopictus and Vero cells was cleaved to produce the membrane-associated virion protein (M) and the non-M fragment (pr) immediately preceding or occurring simultaneously with the release of viral particles from cells. The cleavage was less efficient in mosquito cells. The pr fragment was found only in the medium and was not rapidly degraded. To obtain pr-specific and M-specific antisera for these studies, proteins containing fragments of DEN-2 prM fused with staphylococcal Protein A were synthesized in Escherichia coli using the expression vector pRIT2T. The fusion proteins were stable and were used to raise antisera in rabbits for immunoprecipitation of radiolabelled cell extracts and culture medium. This is the first report of the detection of a C-prM protein in flavivirus-infected cells and the identification of the pr component of prM.
引用
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页码:175 / 182
页数:8
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