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Vectors for Expression and Secretion of FLAG Epitope-Tagged Proteins in Mammalian Cells

Richard G. Chubet

Bill L. Brizzard

Richard G. Chubet

Scientific Imaging Systems, Eastman Kodak Company, New Haven, CT, USA

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Bill L. Brizzard

^*Address correspondence to Bill L. Brizzard, Scientific Imaging Systems, Eastman Kodak Company, 4 Science Park, New Haven, CT 06511, USA. Internet:

E-mail Address: brizzard@ksis.com

Scientific Imaging Systems, Eastman Kodak Company, New Haven, CT, USA

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Published Online:2 Aug 2018https://doi.org/10.2144/96201pf01

Abstract

The FLAG peptide, AspTyrLysAspAspAspAspLys, has been used as an epitope tag in a variety of cell types. The modification of the cytomegalovirus (CMV) promoter containing vector, pCMV5, to create two transient expression vectors designed for secretion and intracellular expression of FLAG-fusion proteins in mammalian cells is described. As a functional test, the bacterial alkaline phosphatase gene was cloned into both vectors, and anti-FLAG monoclonal antibodies were used for detection of FLAG epitope-tagged bacterial alkaline phosphatase in mammalian cells. In addition, secreted bacterial alkaline phosphatase was purified from the extracellular medium by anti-FLAG affinity chromatography.

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Published online 2 August 2018

Published in print January 1996

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