Molecular characterization of Shaker, a Drosophila gene that encodes a potassium channel

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Abstract

The Drosophila Shaker (Sh) gene appears to encode a type of voltage-sensitive potassium (K+) channel called the A channel. We have isolated Sh as part of a 350 kb chromosomal walk. The region around Sh contains four identified transcription units. We find that Sh corresponds to a very large transcription unit encompassing a total of about 95 kb of genomic DNA and split by a major 85 kb intron. Sh has multiple hydrophobic domains that have a high probability of being membrane-spanning, consistent with the proposal that it encodes an ion channel.

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G Rubin, Allan Spradling (1982)

Exogenous DNA sequences were introduced into the Drosophila germ line. A rosy transposon (ry1), constructed by inserting a chromosomal DNA fragment containing the wild-type rosy gene into a P transposable element, transformed germ line cells in 20 to 50 percent of the injected rosy mutant embryos. Transformants contained one or two copies of chromosomally integrated, intact ry1 that were stably inherited in subsequent generations. These transformed flies had wild-type eye color indicating that the visible genetic defect in the host strain could be fully and permanently corrected by the transferred gene. To demonstrate the generality of this approach, a DNA segment that does not confer a recognizable phenotype on recipients was also transferred into germ line chromosomes.

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Transposition of cloned P elements into Drosophila germ line chromosomes.

A. Spradling, G Rubin (1982)

Recombinant DNA carrying the 3-kilobase transposable element was injected into Drosophila embryos of a strain that lacked such elements. Under optimum conditions, half of the surviving embryos showed evidence of P element-induced mutations in a fraction of their progeny. Direct analysis of the DNA of strains derived from such flies showed them to contain from one to five intact 3-kilobase P elements located at a wide variety of chromosomal sites. DNA sequences located outside the P element on the injected DNA were not transferred. Thus P elements can efficiently and selectively transpose from extrachromosomal DNA to the DNA of germ line chromosomes in Drosophila embryos. These observations provide the basis for efficient DNA-mediated gene transfer in Drosophila.