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Journal of Histochemistry and Cytochemistry
Volume 52 (4): 469-477, 2004
Copyright ©The Histochemical Society, Inc.

Production of Cell Lines Secreting TAT Fusion Proteins

Tibor Barka, Edward S. Gresik and Scott C. Henderson

Center for Anatomy and Functional Morphology and Department of Pathology, Mount Sinai School of Medicine, New York, New York (TB); Department of Cell Biology and Anatomical Sciences, City University of New York Medical School, New York, New York (ESG); and Brookdale Department of Molecular Cell and Developmental Biology, Mount Sinai School of Medicine, New York, New York (SCH)

Correspondence to: Tibor Barka, MD, Center for Anatomy and Functional Morphology, Box 1007, Mount Sinai School of Medicine, New York, NY 10029. E-mail: Tibor.Barka{at}mssm.edu

Transduction of proteins and other macromolecules constitutes a potent technology to analyze cell functions and to achieve therapeutic interventions. In general, fusion proteins with protein transduction domains, such as TAT, are produced in a bacterial expression system. Here we describe the generation of a mammalian expression vector coding for TAT-EGFP fusion protein. Transfection of CHO-K1 cells by this vector and subsequent selection by Zeocin resulted in cell lines that express and secrete EGFP, a variant of the green fluorescent protein GFP. The ultimate cell line was produced by first cloning the stable integrants and subsequent selection of EGFP-expressing cells by flow cytometric sorting. In the resulting cell line approximately 98% of cells express EGFP. Using the same methodology, we generated cell lines that express DsRed fluorescent protein. The advantages of using such a mammalian expression system include the ease of generating TAT fusion proteins and the potential for sustained production of such proteins in vitro and, potentially, in vivo. (J Histochem Cytochem 52:469–477, 2004)

Key Words: TAT fusion proteins • green fluorescent protein • red fluorescent protein • mammalian expression vector


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