Expression of Drosophila Shaker potassium channels in mammalian cells infected with recombinant vaccinia virus

Creators: Leonard, R. J.; Karschin, A.; Jayashree-Aiyar, S.; Davidson, N.; Tanouye, M. A.; Thomas, L.; Thomas, G.; Lester, H. A.

Abstract

A recombinant vaccinia virus containing a Drosophila potassium channel (Shaker H4) cDNA was constructed by homologous recombination between wild-type vaccinia virus DNA and a transfer plasmid. The new virus was used to infect four types of mammalian cells in culture. Electrophysiological recording 24-72 hr after infection revealed the expression of voltage-gated transient potassium channels in all four cell types. The properties of the induced currents were identical to those previously observed following injection of the Shaker H4 transcript into oocytes. Vaccinia promises to be an effective vehicle for the heterologous expression of transmembrane ion channels in a variety of cell types.

Additional Information

© 1989 by the National Academy of Sciences. Contributed by N. Davidson, July 14, 1989. We thank Barbara Thorne for expert assistance in the construction of the recombinant virus and J.H. Strauss and R.W. Aldrich for advice. Charybdotoxin was a generous gift of Dr. M.L. Garcia (Merck Sharp & Dohme Research Laboratories). This work was supported by National Institutes of Health Grants GM10991, GM29836, and DK37274, by the Cystic Fibrosis Foundation, and by fellowships from the Max Kade Foundation (A.K.) and the National Institutes of Health (R.J.L.). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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