FUNCTIONAL EXPRESSION OF SHAKER-K+ CHANNELS IN A BACULOVIRUS-INFECTED INSECT CELL-LINE

被引:92
作者
KLAIBER, K
WILLIAMS, N
ROBERTS, TM
PAPAZIAN, DM
JAN, LY
MILLER, C
机构
[1] HARVARD UNIV, SCH MED,DANA FARBER CANC CTR,DEPT PATHOL, DIV CELLULAR & MOLEC BIOL, BOSTON, MA 02115 USA
[2] UNIV CALIF SAN FRANCISCO, MED CTR, HOWARD HUGHES MED INST, DEPT PHYSIOL, SAN FRANCISCO, CA 94143 USA
关键词
D O I
10.1016/0896-6273(90)90311-3
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
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. © 1990.
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页码:221 / 226
页数:6
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