Abstract
Telomeric fragments from salivary gland squashes of Drosophila melanogaster Oregon R. were produced by a new microdissection technique, UV laser microbeam dissection. Microdissection, an essential step in microcloning procedures, is usually performed using micromanipulators and microneedles. Recently it has been shown that microdissection can be improved to very high precision if a laser coupled into a microscope is used. A laser microbeam, generated by an excimer pumped dye laser, allows chromosomes to be cut into slices of less than 0.5 μm. Here it is shown, that single copy DNA probes prepared from Drosophila chromosomes by laser microdissection and microcloning relocalize to the chromosomal regions from which they are derived. The combination of laser technique and microcloning provides an advantageous approach for rapid genetic analysis with potential for the study of genetic diseases and genome mapping.
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Ashkin A, Dziedzic IM, Yamane T (1987) Optical trapping and manipulation of single cells using infrared laser beams. Nature 330:769
Bates GP, Wainwright BJ, Williamson R, Brown SDM (1986) Microdissection of and microcloning from the short arm of human chromosome 2. Mol Cell Biol 6:3826–3830
Berns MW, Aist J, Edwards J, Strahs K, Girton J, McNeil P, Rattner JB, Kitzes M, Hammer-Wilson M, Liaw L-H, Siemens A, Koonce M, Peterson S, Brenner S, Burt J, Walter R, Bryant PJ, van Dyk D, Coulombe J, Cahill T, Berns GS (1981) Laser microsurgery in cell and developmental biology. Science 213:505–513
Brockdorff N, Fisher EMC, Cavanna JS, Lyon MF, Brown SDM (1987) Construction of a detailed molecular map of the mouse X chromosome by microcloning and interspecific crosses. EMBO J 6:3291–3297
Calmettes PP, Berns MW (1983) Laser induced multiphoton processes in living cells. Proc Natl Acad Sci USA 80:7197–7199
Collins FS, Weissmann SM (1984) Directional cloning of DNA fragments at a large distance from an initial probe: A circularization method. Proc Natl Acad Sci USA 81:6812–6816
Cormona MJ, Morcillo G, Galler R, Martinez-Salas E, dela Campa AG, Diez JL, Edström JE (1985) Cloning and molecular characterization of a telomeric sequence from a temperature induced Balbiani ring. Chromosoma 92:108–115
Cross SH, Little PFR (1986) A cosmid vector for systematic chromosome walking. Gene 49:9–22
Fisher EMC, Cavanna JS, Brown SDM (1985) Microdissection and microcloning of the mouse X chromosome. Proc Natl Acad Sci USA 82:5846–5849
Greulich KO, Monajembashi S, Cremer T, Cremer C, Butz P, Wolfrum J (1986) Micromanipulation of biological cells and chromosomes by an excimer laser pumped dye microbeam. International Quantum Electronics Conference, San Francisco, paper TuGG11
Hennig W, Huijser P, Vogt P, Jäckle H, Edström JE (1983) Molecular cloning of microdissected lampbrush loop DNA sequences of Drosophila hydei. EMBO J 2:1741–1746
Hohn B (1979) In vitro packaging of lambda and cosmid DNA. Methods Enzymol 68:299
Kaiser R, Weber J, Grzeschik K-H, Edström JE, Driesel A, Zengerling S, Buchwald M, Tsui C, Olek K (1987) Microdissection and microcloning of the long arm of human chromosome 7. Mol Biol Rep 12:3–6
Lathe R, Lecocq JP (1977) Overproduction of a viral protein during infection of a lyc mutant of Escherichia coli with phage lambda imm 434. Virology 83:204–206
Maniatis T, Fritsch EF, Sambrock J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, New York
Monajembashi S, Cremer C, Cremer T, Wolfrum J, Greulich KO (1986) Microdissection of human chromosomes by a laser microbeam. Exp Cell Res 167:262–265
Murray NE, Brammer WJ, Murray K (1977) Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet 150:53–61
Pardue ML, Gall JG (1975) Nuclei acid hybridization to the DNA of cytological preparation. Methods Cell Biol 10:1–16
Pirrotta V, Jäckle H, Edström JE (1983a) Genetic engineering: Principles and methods, vol 5. Setlow JK, Hollaender A (eds) Plenum, New York London, p 1–17
Pirrotta V, Hadfield C, Pretorius GHJ (1983b) Microdissection and cloning of the white locus and the 3B1-3C2 region of the Drosophila X chromosome. EMBO J 2:927–934
Preiss A, Rosenberg Urs B, Kienlin A, Seifert E, Jäckle H (1985) Molecular genetics of Krüppel, a gene required for segmentation of the Drosophila embryo. Nature 313:27–32
Renkawitz-Pohl R, Glätzer KH, Kunz W (1980) Characterization of cloned ribosomal DNA from Drosophila hydei. Nucleic Acids Res 8:4593–4611
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Röhme D, Fox H, Herrmann B, Frischauf AM, Edström JE, Mains P, Silver LM, Lehrach H (1984) Molecular clones of the mouse t complex derived from microdissected metaphase chromosomes. Cell 36:783–788
Scalenghe F, Turco E, Edström JE, Pirrotta V (1981) Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes. Chromosoma 82:205–216
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Staller Young B, Pession A, Traverse KL, French C, Pardue ML (1983) Telomere regions in Drosophila share complex DNA sequences with pericentric heterochromatin. Cell 34:85–94
Steinemann M (1984) Telomere repeats within the neo-Y-chromosome of Drosophila miranda. Chromosoma 90:1–5
Wiegand R, Weber G, Zimmermann K, Monajembashi S, Wolfrum J, Greulich KO (1987) Laser-induced fusion of mammalian cells and plant protoplasts. J Cell Sc 88:145–149
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Ponelies, N., Bautz, E.K.F., Monajembashi, S. et al. Telomeric sequences derived from laser-microdissected polytene chromosomes. Chromosoma 98, 351–357 (1989). https://doi.org/10.1007/BF00292388
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DOI: https://doi.org/10.1007/BF00292388