Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P element transposon tagging

Cell. 1982 Dec;31(3 Pt 2):585-92. doi: 10.1016/0092-8674(82)90314-2.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles
  • Animals
  • Cloning, Molecular*
  • DNA Restriction Enzymes
  • DNA Transposable Elements*
  • DNA-Directed RNA Polymerases / genetics*
  • Drosophila melanogaster / enzymology*
  • Drosophila melanogaster / genetics
  • Female
  • Genes*
  • Genes, Lethal
  • Genes, Recessive
  • Mutation
  • Nucleic Acid Hybridization
  • Protein Biosynthesis
  • RNA Polymerase II / genetics*

Substances

  • DNA Transposable Elements
  • RNA Polymerase II
  • DNA-Directed RNA Polymerases
  • DNA Restriction Enzymes