Localizing genes in Drosophila melanogaster polytene chromosomes by fluorescence in situ hybridization.
This paper describes a method for the identification of single copy genes in Drosophila melanogaster polytene chromosomes, using fluorescence in situ hybridization (FISH). We demonstrate the detection of white (w), a gene previously mapped to 1-1.5 region of the linkage map, and to 3C2 region of the cytogenetic map of X chromosome. Squash preparations of polytene chromosomes from salivary glands dissected out from third instar larvae of Drosophila melanogaster were denatured and subjected to hybridization with a digoxigenin labeled probe, corresponding to mini-white gene. The preparations were then washed and incubated with anti-digoxigenin-fluorescein antibodies. After removal of the nonspecifically bound antibodies, the polytene chromosomes were counterstained with propidium iodide. Fluorescence microscopy revealed white locus in the X chromosome in a subterminal location, in agreement with the above mentioned maps. The protocol is efficient and adaptable for simultaneously multiple signal detection.
Gavrila
L
L
Department of Genetics, Faculty of Biology, University of Bucharest, CENTRAGEN, 1-3 Portocalelor, 76258, sect.6, Bucharest, Romania. gavrila@botanic.unibuc.ro
Ecovoiu
A A
AA
Georgescu
L M
LM
D018528:ATP-Binding Cassette Transporters; D000818:Animals; D002875:Chromosomes / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics* / Q000254:growth & development; D005136:Eye Proteins / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D020161:Physical Chromosome Mapping
D018528:ATP-Binding Cassette Transporters; D029721:Drosophila Proteins; D005136:Eye Proteins; C059254:w protein, Drosophila