Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes

Fragments from section 3 of the salivary gland X chromosome of D. melanogaster were dissected with a micromanipulator. The DNA was extracted, cut and ligated to a λ vector in a volume of a few nanoliters in an oil chamber monitored through a microscope. From about 10 pg of DNA we obtained 80 recombinant clones, a sample of which were analysed and shown to contain Drosophila DNA which hybridises in situ to the region of section 3 of the X chromosome. With this technique we can isolate clones from any desired region as small as 200 kb from the euchromatic arms of polytene chromosomes.

[1]  H. Goodman,et al.  A restriction endonuclease analysis of the bacterial plasmid controlling the ecoRI restriction and modification of DNA. , 1976, Federation proceedings.

[2]  K. Kaiser,et al.  Physical characterisation of the “Rac prophage” in E. coli K12 , 1979, Molecular and General Genetics MGG.

[3]  D. Poulson.  Chromosomal Control of Embryogenesis in Drosophila , 1945, The American Naturalist.

[4]  C. Bridges.  A REVISED MAP OF THE SALIVARY GLAND X-CHROMOSOME of Drosophila Melanogaster , 1938 .

[5]  E. Southern.  Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[6]  J. Gall,et al.  Nucleic acid hybridization to the DNA of cytological preparations. , 1975, Methods in cell biology.

[7]  L. Enquist,et al.  In vitro packaging of a lambda Dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1. , 1977, Gene.

[8]  G. Lefevre.  photographic representation and interpretation of the polytene chromosomes of Drosophila melanogaster salivary glands , 1976 .

[9]  T. Kaufman,et al.  The anatomy and function of a segment of the X chromosome of Drosophila melanogaster. , 1972, Genetics.

[10]  Calvin B. Bridges,et al.  SALIVARY CHROMOSOME MAPSWith a Key to the Banding of the Chromosomes of Drosophila Melanogaster , 1935 .

[11]  L. Enquist,et al.  In vitro packaging of a λ Dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1 , 1977 .

[12]  B. Judd,et al.  Allelic pairing and gene regulation: A model for the zeste-white interaction in Drosophila melanogaster. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[13]  G. Lefevre.  The relationship between genes and polytene chromosome bands. , 1974, Annual review of genetics.

[14]  David M. Prescott,et al.  Methods in cell physiology , 1964 .

[15]  R. Goldschmidt.  Chromosomes and genes. , 1951, Cold Spring Harbor symposia on quantitative biology.

[16]  W. J. Brammar,et al.  Lambdoid phages that simplify the recovery of in vitro recombinants , 1977, Molecular and General Genetics MGG.

[17]  W. Beermann.  Chromomeres and genes. , 1972, Results and problems in cell differentiation.

[18]  H. L. Carson,et al.  The Genetics and Biology of Drosophila , 1976, Heredity.

[19]  J. Edström.  Chapter 22 Microextraction and Microelectrophoresis for Determination and Analysis of Nucleic Acids in Isolated Cellular Units , 1964 .

[20]  F. Blattner,et al.  Cloning human fetal gamma globin and mouse alpha-type globin DNA: preparation and screening of shotgun collections. , 1978, Science.

[21]  B. Hohn,et al.  Packaging recombinant DNA molecules into bacteriophage particles in vitro. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[22]  D T Denhardt,et al.  A membrane-filter technique for the detection of complementary DNA. , 1966, Biochemical and biophysical research communications.

[23]  F. Blattner,et al.  Cloning human fetal gamma globin and mouse alpha-type globin DNA: characterization and partial sequencing. , 1978, Science.

[24]  R. Hardison,et al.  The isolation of structural genes from libraries of eucaryotic DNA , 1978, Cell.