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Neuron
Article . 1990 . Peer-reviewed
License: Elsevier TDM
Data sources: Crossref
Neuron
Article . 1990
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Functional expression of Shaker K+ channels in a baculovirus-infected insect cell line

Authors: Lily Yeh Jan; Thomas M. Roberts; Kimberly Klaiber; Christopher Miller; Diane M. Papazian; Nidhi Gupta Williams;

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.

Related Organizations
Keywords

Potassium Channels, Base Sequence, Blotting, Western, Molecular Sequence Data, Electric Conductivity, Insect Viruses, DNA, Moths, Cell Line, Kinetics, Gene Expression Regulation, Animals, Drosophila, Cloning, Molecular, Promoter Regions, Genetic

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    102
    popularity
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Powered by OpenAIRE graph
citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
102
Average
Top 1%
Top 1%