Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P element transposon tagging.
We have identified a lethal mutation in the D. melanogaster RNA polymerase II locus, RpIIC4, caused by insertion of a transposable element associated with the phenomenon of hybrid dysgenesis (P element). Using previously cloned P element sequences as a hybridization probe we have isolated a hybrid lambda phage clone carrying a 10 kb genomic DNA fragment containing a 1.3 kb P element insert and flanking sequences from the RpII locus. The non-P sequences in this clone (lambda DmRpII-1) hybridize to polytene chromosome band region 10C, the cytogenetic location of RpIIC4, and revertants which lose the lethal RNA polymerase II mutation also lose P element sequences from the locus. We have generated several additional P element insertions into the locus and shown that they can occur at two or more different sites. These experiments illustrate that mutagenesis by P element insertion and use of cloned P DNA to retrieve the DNA sequences into which insertion has occurred may be a general method for cloning genetically defined loci in Drosophila.
D000483:Alleles; D000818:Animals; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D005260:Female; D005796:Genes; D005804:Genes, Lethal; D005808:Genes, Recessive; D009154:Mutation; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012319:RNA Polymerase II / Q000235:genetics*
D004251:DNA Transposable Elements; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; D004262:DNA Restriction Enzymes