FOOT-AND-MOUTH-DISEASE VIRUS 2A OLIGOPEPTIDE MEDIATED CLEAVAGE OF AN ARTIFICIAL POLYPROTEIN

被引:308
作者
RYAN, MD
DREW, J
机构
[1] AFRC Institute for Animal Health, Pirbright Laboratory, Pirbright, Woking, Surrey GU24 0NF, Ash Road
关键词
AUTOPROTEOLYTIC; COTRANSLATIONAL CLEAVAGE; FOOT-AND-MOUTH DISEASE VIRUS; POLYPROTEIN;
D O I
10.1002/j.1460-2075.1994.tb06337.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We describe the construction of a plasmid (pCAT2AGUS) encoding a polyprotein in which a 19 amino acid sequence spanning the 2A region of the foot-and-mouth disease virus (FMDV) polyprotein was inserted between the reporter genes chloramphenicol acetyl transferase (CAT) and beta-glucuronidase (GUS) maintaining a single, long open reading frame. Analysis of translation reactions programmed by this construct showed that the inserted FMDV sequence functioned in a manner similar to that observed in FMDV polyprotein processing: the CAT2AGUS polyprotein underwent a cotranslational, apparently autoproteolytic, cleavage yielding CAT-2A and GUS. Analysis of translation products derived from a series of constructs in which sequences were progressively deleted from the N-terminal region of the FMDV 2A insertion showed that cleavage required a minimum of 13 residues. The FMDV 2A sequence therefore provides the opportunity to engineer either whole proteins or domains such that they are cleaved apart cotranslationally with high efficiency.
引用
收藏
页码:928 / 933
页数:6
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