UGUS, a reporter for use with destabilizing N-termini

被引:5
作者
Zaccaria, D
Greco, R
MacWilliams, H
Bozzaro, S
Ceccarelli, A [1 ]
机构
[1] Univ Turin, Osped S Luigi, Dipartimento Sci Clin & Biol, I-10043 Turin, Italy
[2] Univ Munich, Inst Zool, D-80333 Munich, Germany
关键词
D O I
10.1093/nar/26.4.1128
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
Due to constraints in vector construction, reporter polypeptides often carry N-terminal sequences of extraneous origin, Since protein half-life can be influenced by small determinants in the N-terminus, such foreign sequences can destabilize proteins and compromise results of reporter-based studies, We provide a real-life example of this problem (destabilizing sequences derived from a ribosomal protein) and show that it can be solved with the ubiquitin fusion technique, in which ubiquitin sequences are placed upstream of the reporter, in our case beta-glucuronidase. Post-translational processing by characterized pathways removes the ubiquitin together with destabilizing sequences, generating a stable reporter whose N-terminus is constant.
引用
收藏
页码:1128 / 1129
页数:2
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