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Detection of in situ cleaved p115 with the cut specific antibodies in rapid protein inactivation system by tobacco etch viral protease cleavage

Authors Koreishi, Honjo, Satoh A

Published 10 August 2011 Volume 2011:1 Pages 5—11


Review by Single-blind

Peer reviewer comments 4

Mayuko Koreishi1,2, Yasuko Honjo1, Ayano Satoh1,2
1The Research Core for Interdisciplinary Sciences (RCIS), Okayama University, Okayama, Japan; 2Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan

Abstract: Gene perturbation methods are commonly used in the study of gene and protein function. The authors of this paper recently developed a rapid protein inactivation technique utilizing tobacco etch virus (TEV)-derived protease. TEV protease recognizes the ENLYFQG (Glu-Asn-Leu-Tyr-Phe-Gln-Gly) amino acid sequence and specifically cleaves between Q and G. The authors developed antibodies that recognize the cleaved TEV (ENLYFQ) sequence, both in vitro and in vivo, but do not bind to uncleaved TEV (ENLYFQG). Using these antibodies, in situ protein cleavage was successfully detected. These antibodies used in combination with the TEV protease may be a useful complement to other perturbation methods.

Keywords: TEV protease, Golgi, golgins, microinjection, recombinant proteins

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