Electron microscopic in situ hybridization of digoxigenin-dUTP-labelled DNA probes with Drosophila melanogaster polytene chromosomes.
We report a simplified method of electron microscopic (EM) in situ hybridization for standard squashes of Drosophila melanogaster polytene chromosomes using digoxigenin-11-dUTP labelled DNA probes. The method is efficient and reproducible: its high resolution and specificity were demonstrated for the transformed strain 148, in which the insertion was localized precisely as a new thin band both by conventional EM and according to our method. In addition, the method was applied to the fine mapping of the developmentally regulated gene muscle-blind (mbl). On the one hand, mbl was shown to cover the 54B1-2 large band and the adjacent interbands in the 2R polytene chromosome. On the other hand, the use of distantly located DNA probes in the mbl gene allowed us to orientate the transcription unit in the chromosome.
Semeshin
V F
VF
Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk. semeshin@bionet.nsc.ru
Artero
R
R
Perez-Alonso
M
M
Shloma
V V
VV
D000818:Animals; D002874:Chromosome Mapping; D002875:Chromosomes; D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D004076:Digoxigenin / Q000031:analogs & derivatives*; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics*; D017403:In Situ Hybridization / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods; D009687:Nuclear Proteins / Q000235:genetics
D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D029721:Drosophila Proteins; D009687:Nuclear Proteins; C070202:digoxigenin-11-deoxyuridine triphosphate; C109271:mbl protein, Drosophila; D004076:Digoxigenin