EXPRESSION IN ESCHERICHIA-COLI AND PURIFICATION OF BIOLOGICALLY-ACTIVE L-PROTEINASE OF FOOT-AND-MOUTH-DISEASE VIRUS

被引：15

作者：

PICCONE, ME ^{[1
]}

SIRA, S ^{[1
]}

ZELLNER, M ^{[1
]}

GRUBMAN, MJ ^{[1
]}

机构：

[1] USDA ARS,PLUM ISL ANIM DIS CTR,GREENPORT,NY 11944

来源：

VIRUS RESEARCH | 1995年 / 35卷 / 03期

关键词：

FOOT-AND-MOUTH DISEASE VIRUS; LEADER PROTEINASE; AUTOCATALYTIC CLEAVAGE; P220; CLEAVAGE;

D O I：

10.1016/0168-1702(94)00084-P

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The foot-and-mouth disease virus (FMDV) Lb gene was cloned into bacterial expression vectors under the control of a T7 RNA polymerase promoter. The Lb protein was expressed in both an in vitro transcription-translation system and in Escherichia coli. In vitro expression of a construct containing the Lb gene fused to a portion of the VP4 and 3D genes demonstrated cis cleavage activity that could be blocked by the thiol protease inhibitor E-64. Lb expressed in E. coli was purified from the soluble fraction by metal chelation chromatography. Purified Lb had trans cleavage activity at the L/P1 junction and cleaved the p220 component of the cap-binding protein complex.

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页码：263 / 275

页数：13

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