IN-VITRO ASSEMBLY OF COWPEA CHLOROTIC MOTTLE VIRUS FROM COAT PROTEIN EXPRESSED IN ESCHERICHIA-COLI AND IN VITRO-TRANSCRIBED VIRAL CDNA
IN-VITRO ASSEMBLY OF COWPEA CHLOROTIC MOTTLE VIRUS FROM COAT PROTEIN EXPRESSED IN ESCHERICHIA-COLI AND IN VITRO-TRANSCRIBED VIRAL CDNA
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DOI：
10.1006/viro.1995.1108
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发表时间：
1995-03-10
期刊：
影响因子：
3.7
通讯作者：
YOUNG, MJ
中科院分区：
文献类型：
--
作者：
ZHAO, XX;FOX, JM;YOUNG, MJ
小球形植物病毒--豌豆褪绿斑驳病毒(CCMV)为检测球形病毒组装提供了一种理想的系统。我们对CCMV的体外组装系统进行了改进，使其能够从在大肠杆菌中表达的外壳蛋白和从全长cDNA体外转录的病毒RNA中产生病毒粒子。用低温电子显微镜和图像重建技术对体外组装的颗粒进行检测表明，在2.5 nm分辨率下，这些颗粒与植物纯化的颗粒是无法区分的。对外壳蛋白N-端和C-端延伸的突变分析表明，它们在病毒组装中各自发挥作用。N-末端是组装含有颗粒的RNA所必需的，而不是组装空病毒粒子所必需的。C末端对于外壳蛋白二聚体的形成和颗粒组装是必不可少的。(C)1995年学术出版社。
The small spherical plant virus, cowpea chlorotic mottle virus (CCMV), provides an ideal system to examine spherical virus assembly. We have modified the CCMV in vitro assembly system to produce virions from coat protein expressed in Escherichia coli and viral RNA transcribed in vitro from full-length cDNAs. Examination of the in vitro-assembled particles with cryoelectron microscopy and image reconstruction techniques demonstrates that the particles are indistinguishable from plant purified particles at 2.5 nm resolution. Mutational analysis of the coat protein N- and C-terminal extensions demonstrate their respective roles in virus assembly. The N-terminus is required for assembly of RNA containing particles but not for the assembly of empty virions. The C-terminus is essential for coat protein dimer formation and particle assembly. (C) 1995 Academic Press, Inc.