Open AccessMolecular cloning of a Drosophila potential Z-DNA forming sequence hybridizingin situto a developmentally regulated subdivision of the polytene chromosomes
Author(s) -
Antonio Jiménez-Ruı́z,
José Marı́a Requena,
Francesca Lancillotti,
Glauco Morales,
Manuel Carlos López López,
Carlos Alonso
Publication year - 1989
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/17.12.4579
Subject(s) - biology , polytene chromosome , plasmid , insert (composites) , microbiology and biotechnology , dna , molecular cloning , recombinant dna , genetics , locus (genetics) , in vitro recombination , drosophila melanogaster , gene , peptide sequence , mechanical engineering , engineering
We describe the selection of a group of plasmids with potential to form Z-DNA, from libraries of Drosophila hydei nuclear DNA using anti Z-DNA monoclonal (22) or polyclonal (10c) antibodies. The supercoiled closed circular forms of most of the selected recombinant plasmids from the 10c Z-DNA library show affinity to the polyclonal 10c antibody as indicated by DNA binding assays. One of these plasmids, pF17, was selected for further study. The insert in this plasmid adopts the Z conformation at bacterial supercoiled density. Analysis of deletion plasmids indicates that a Z-epitope is located within a short fragment of the insert in which 3 GC repetitions are found. The Drosophila DNA insert in pF17 hybridizes in situ with locus 4-75C1-2 of the polytene chromosomes, a locus whose transcriptional activity is developmentally regulated during the third instar.