A functional hepatitis B virus X protein produced in insect cells.
A functional hepatitis B virus X protein produced in insect cells.
复制标题
昆虫细胞中产生的功能性乙型肝炎病毒 X 蛋白。
DOI：
10.1016/0042-6822(91)90577-x
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发表时间：
1991
期刊：
影响因子：
3.7
通讯作者：
Lee,CH
中科院分区：
文献类型：
--
作者：
Spandau,DF;Wang,HG;Fraser,MJ;Lee,CH
We have constructed a recombinant baculovirus fromAutographa californicanuclear polyhedrosis virus, called AcX, that expresses the gene encoding the hepatitis B virus X protein in infectedSpodoptera frugiperda(Sf21AE) insect cells. A 16.5-kDa monomer and a 33-kDa dimer of the X protein were detected in extracts from AcX-infected cells on immunoblots using a polyclonal anti-X antibody. The biological activity of the insect cell-produced X protein was assayed by fusing AcX-infected Sf21 AE cells with African green monkey kidney (CV-1) cells and then transfecting the fused cells with the reporter plasmid pSV2cat. The expression of the cat gene in CV- 1:Sf21AE(AcX) fusions was seven times higher than that derived from CV-1:Sf21AE(AcMNPV) fusions, indicating that the insect cell-produced X protein was functional. The transactivation function of the insect cell-produced X protein was also verified by scrape-loading nuclear extracts of AcX-infected Sf21AE cells into pSV2cat-transfected CV-1 cells. Treatment of the AcX-infected cell nuclear extracts with anti-X antisera prior to scrape-loading eliminated the transactivating activity of the extracts. We conclude that the insect cell-produced X protein was functionally identical to that generated in mammalian cells.
