Functional expression of Shaker K+ channels in a baculovirus-infected insect cell line.

Abstract:

:We constructed a recombinant baculovirus, A. californica nuclear polyhedrosis virus, containing the Drosophila Shaker H4 K+ channel cDNA under control of the polyhedrin promoter. When infected with this recombinant baculovirus, the cell line Sf9, derived from the army-worm caterpillar S. frugiperda, expresses fully functional Shaker transient K+ currents, as assayed by whole-cell recording. K+ currents begin to appear at about 15 hr after infection, and they continue to increase over the next 3 days. Over the same period of time, a 75 kd band appears on SDS gels stained with Coomassie blue. The identity of this band as a Shaker gene product is confirmed by Western blot analysis using an anti-Shaker antiserum. The 75 kd band accounts for a substantial fraction of the membrane protein in Shaker-infected Sf9 cells. These results give hope that the baculovirus system, which has been used successfully for high-level expression of soluble proteins from higher eukaryotes, may be appropriate for producing large amounts of cloned ion channel proteins as well.

journal_name

Neuron

journal_title

Neuron

authors

Klaiber K,Williams N,Roberts TM,Papazian DM,Jan LY,Miller C

doi

10.1016/0896-6273(90)90311-3

subject

Has Abstract

pub_date

1990-08-01 00:00:00

pages

221-6

issue

2

eissn

0896-6273

issn

1097-4199

pii

0896-6273(90)90311-3

journal_volume

5

pub_type

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