We have cloned vermilion (v), one of the genes required for brown eye pigment synthesis in Drosophila, using a mutant (vH2a) "tagged" with the transposon P factor.Mutations that disrupt v gene expression are clustered within approximately 2 kilobases of DNA. A 1.4-kilobase transcript, homologous to this same region, is present in v+ RNA but absent in RNA from several v mutants. The spontaneous v alleles that are suppressed by the suppressor of sable [su(s)
MATERIALS AND METHODSPurification of Nucleic Acids. The v mutants used for nucleic acid isolation are described by Lindsley and Grell (16) or are referenced in the text. Drosophila DNA was isolated from adult flies as described by Bingham et al. (18). Procedures for isolation ofbacteriophage X DNA and plasmid DNA have been described (19). Total RNA was prepared from 2-day-old adults by using procedures described by Thireos et al. (20) except that 10 mM vanadyl-ribonucleoside complex was included in the homogenization buffer. Poly(A)+ RNA was purified by chromatography on oligo(dT)-cellulose (21 nitrocellulose (26). The hybridization conditions were the same as described for Southern transfers. The hybridization probes were plasmid subclones that had been labeled with 32P by nick-translation (27).Abbreviations: kb, kilobase(s); bp, base pair(s).
404The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.