FUNCTIONAL EXPRESSION OF SHAKER-K+ CHANNELS IN A BACULOVIRUS-INFECTED INSECT CELL-LINE
FUNCTIONAL EXPRESSION OF SHAKER-K+ CHANNELS IN A BACULOVIRUS-INFECTED INSECT CELL-LINE
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DOI：
10.1016/0896-6273(90)90311-3
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发表时间：
1990-08-01
期刊：
影响因子：
16.2
通讯作者：
MILLER, C
中科院分区：
文献类型：
--
作者：
KLAIBER, K;WILLIAMS, N;MILLER, C
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 sllbstantial 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.
