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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.
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页码:928 / 933
页数:6
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