Abstract
We have isolated by molecular cloning techniques a structural gene that maps to subregion 63 F of Drosophila melanogaster chromosome 3L. This locus being one of the early chromosomal targets for ecdysone stimulation, may be induced to puff by the hormone. The gene is on a plasmid vector and it has been designated as pDm 63F. This recombinant molecule also maps to the early ecdysone inducible subregion 90B of Drosophila hydei chromosome 4. The cytological inspection of large number of chromosomal sets after in situ hybridization of the cloned DNA, locates the cloned sequence between bands 63F 2–4 according to Bridges map. Similarly, in Drosophila hydei the cloned DNA maps between subdivisions 90B 2–4 according to Berendes’ map. In situ hybridization of the pDm 63F cloned DNA, directed to the nascent RNA rather than to the DNA, shows two to three times more silver grains over the corresponding regions when puffed than in the resting stage. There are however quantitative differences attending to the transcriptional activity of homologous loci in both species. By RNA excess hybridization we have found that the cellular concentration of the 63F cloned mRNA is about 2 times higher in hormone stimulated total larval tissues than in non-stimulated ones.
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Alonso, C., Barberá, E., Pagés, M.: Absolute DNA content of polytene nuclei of D.hydei. Dros. Inf. Serv. 53, 149 (1978)
Ashburner, M.: Puffing patterns in Drosophila melanogaster and related species. In: Developmental studies on giant chromosomes (W. Beermann, ed.), vol. 4, pp. 101–151. Berlin, Heidelberg, New York: Springer, 1972
Ashburner, M.: Sequential gene activation by ecdysone in polytene chromosomes of Drosophila melanogaster I. Dependence upon ecdysone concentrations. Dévelop. Biol. 35, 47–61 (1973)
Ashburner, M.: Chromosomal action of ecdysone. Nature (Lond.) 285, 435–436 (1980)
Becker, H.J.: Die Puffs der Speicheldrüsenchromosomen von Drosophila melanogaster. II. Mitteilung. Chromosoma (Berl.) 13, 341–384 (1962)
Beermann, W.: Chromosomeres and genes. In: Developmental Studies on giant chromosomes (W. Beermann, ed.), vol. 4, pp. 1–33. Berlin, Heidelberg, New York: Springer 1972
Belyaeva, E.S., Aizenzon, M.G., Semeshin, V.F., Kiss, I.I., Koczka, K., Baritcheva, E.M., Gorolova, T.D., Zhimulev, I.F.: Cytogenetic analysis of the 2B 3-4-2 B 11 region of the X-chromosome of Drosophila melanogaster. Chromosoma (Berl.) 81, 281–306 (1980)
Berendes, H.D.: The salivary gland chromosomes of Drosophila hydei Sturtevant. Chromosoma (Berl.) 14, 195–206 (1963)
Berendes, H.D.: The induction of changes in chromosomal activity in different polytene types of cell in Drosophila hydei. Develop. Biol. 11, 371–384 (1965)
Berendes, H.D.: The hormone ecdysone as effector of specific changes in the pattern of gene activities of Drosophila hydei. Chromosoma (Berl.) 22, 274–293 (1967)
Berendes, H.D., Alonso, C., Helmsing, P.J., Leenders, H.J., Derksen, J.: Structure and function in the genome of Drosophila hydei. Cold Spr. Harb. Symp. quant. Biol. 37, 645–654 (1973)
Borst, D.W., Bollenbacher, W.E., O'Connor, J.D., Fristrom, J.W.: Ecdysone levels during metamorphosis of Drosophila melanogaster. Develop. Biol. 39, 308–316 (1974)
Bridges, C.B.: Salivary chromosome maps with a key to the banding of the chromosomes of Drosophila melanogaster. J. Hered. 26, 60–64 (1935)
Casey, J., Davidson, N.: Rates of formation and thermal stabilities of RNA: DNA and DNA: RNA duplexes at high concentrations of formamide. Nucleic Acids Res. 4 1539–1552 (1977)
Clarke, L., Carbon, J.: A colony bank containing synthetic coli E. hybrid plasmids representative of the entire E. coli genome. Cell 9, 91–99 (1976)
Davidson, E.H., Britten, R.J.: Regulation of gene expression possible role of repetitive sequences. Science 204, 1052–1059 (1979)
Gergen, J.P., Stern, R.H., Wensink, P.C.: Filter replicas and permanent collection of recombinant DNA plasmids. Nucleic Acids. Res. 7, 2119–2136 (1979)
Hastie, N.D., Bishop, J.O.: The expression of three abundance clases of mRNA in mouse tissues. Cell 9, 761–771 (1976)
Izquierdo, M., Bishop, J.: An analysis of cytoplasmic RNA populations in Drosophila melanogaster Oregon R. Biochem. Genet. 17, 473–497 (1979)
Judd, B.H., Shen, M.W., Kaufman, T.C.: The anatomy and function of a segment of the X-chromosome of Drosophila melanogaster. Genetics 71, 139–156 (1972)
Keppy, D.O., Welshons, W.J.: The synthesis of compound bands in Drosophila melanogaster salivary gland chromosomes. Chromosoma (Berl.) 76, 191–200 (1980)
Laird, C.D.: DNA of Drosophila chromosomes. Ann. Rev. Genet. 7, 177–204 (1973)
Laird, C.D.: Structural paradox of polytene chromosomes. Cell 22, 869–974 (1980)
Lefevre, G. Jr.: The relationship between genes and polytene chromosome bands. Ann. Rev. Genet. 8 51–62 (1974)
Livak, K.J., Freund, R., Scheweber, M., Wensink, P.C., Meselson, M.: Sequence organization and transcription at two heat shock loci in Drosophila. Proc. nat. Acad. Sci. (Wash.) 75, 5613–5617 (1978)
Maizels, N.: Dyctoystelium 17S, 25S and 5S rDNAs lie within a 38,000 base pairs repeated unit. Cell 9, 431–438 (1976)
Maniatis, T., Jeffrey, A., Van de Sande, H.: Chain length determination of small double and single-stranded DNA molecules by polycrylamyde gel electrophoresis. Biochemistry 14, 3787–3794 (1975)
Pages, M., Alonso, C.: Chemical and conformational changes in chromosome regions being actively transcribed. Nucleic Acids Res. 5, 549–562 (1978)
Patterson, J.T., Stone, W.S.: Evolution in the genus Drosophila. New York: Macmillan. Co. 1952
Richards, G.: Ecdysteroids and puffing in Drosophila melanogaster. In: Progress in ecdysone research (J.A. Huffman, ed.), pp. 363–378. Amsterdam: Elsevier, North-Holland Biomedica Press, 1980
Rigby, P.W.J., Dieckmann, M., Rhodes, C., Berg, P.: Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. molec. Biol. 113, 237–251 (1977)
Ross, J., Aviv, M., Scolnick, E., Leder, P.: In vitro synthesis of DNA complementary to purified rabbit globin mRNA. Proc. nat. Acad. Sci. (Wash.) 69, 264 (1972)
Rudkin, G.T.: Replication in polytene chromosomes. In: Developmental studies on giant chromosomes (W. Beermann, ed.), vol. 4, pp. 58–85. Berlin, Heidelberg, New York: Springer 1972
Rudkin, G.T., Stollar, B.D.: High-resolution detection of DNA-RNA hybrids in situ by indirect inmunofluorescence. Nature (Lond.) 265, 472–473 (1977)
Weintraub, H.: Recognition of specific DNA sequences in eukaryotic chromosomes. Nucleic Acids Res. 8, 4745–4753 (1980)
Zhimulev, I.F., Belyaeva, E.S.: Proposals to the problem of structural and functional organization of polytene chromosomes. Theor. appl. Genet. 45, 335–340 (1975)
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The authors dedicate this paper to the memory of Hans D. Berendes whose work provided some of the fundaments for the present research
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Izquierdo, M., Arribas, C. & Alonso, C. Isolation of a structural gene mapping to subregions 63F of Drosophila melanogaster and 90B of D. hydei polytene chromosomes. Chromosoma 83, 353–366 (1981). https://doi.org/10.1007/BF00327358
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DOI: https://doi.org/10.1007/BF00327358