Mapping the domains on the phosphoprotein of bovine respiratory syncytial virus required for N-P interaction using a two-hybrid system

被引:24
作者
Mallipeddi, SK [1 ]
Lupiani, B [1 ]
Samal, SK [1 ]
机构
[1] UNIV MARYLAND,VIRGINIA MARYLAND REG COLL VET MED,COLLEGE PK,MD 20742
关键词
D O I
10.1099/0022-1317-77-5-1019
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
Specific interactions between the nucleocapsid protein (N) and the phosphoprotein (P) of bovine respiratory syncytial virus (BRSV) have been investigated using a yeast-based two-hybrid system. Plasmids encoding the yeast GAL4 DNA binding domain fused with the N gene and GAL4 activation domain fused with the P gene were contransfected into competent yeast cells. The ability of the N and P proteins to interact in vivo was measured by activation of the lacZ reporter gene by the GAL4 transactivation region. Results indicated that the N and P protein interact very strongly in vivo. When interactions between N and various deletion mutants of the P protein were examined, an internal region (aa 132-168) and the highly acidic C-terminal region (aa 236-241) of the P protein were found to be essential for N-P interaction. In addition, the highly basic N-terminal region (amino acids 1-40) was found to be involved in N-P interaction to a lesser extent.
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页码:1019 / 1023
页数:5
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