Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1984 Sep;3(9):2077–2085. doi: 10.1002/j.1460-2075.1984.tb02094.x

Functional analysis of the white+ gene of Drosophila by P-factor-mediated transformation

Walter J Gehring 1, Roman Klemenz 1, Ursula Weber 1, Urs Kloter 1
PMCID: PMC557646  PMID: 16453549

Abstract

A 12-kb DNA segment spanning the white+ (w+) locus of Drosophila has been inserted into a P-transposon vector and used for P-factor-mediated germ-line transformation. Several red-eyed transformants were recovered which complement the white mutant phenotype. Analysis of the eye pigments and the interaction with the zeste mutation indicates that the w+ gene inserted at several new chromosomal sites is expressed normally. The tissue-specific accumulation of w+ transcripts, as studied by in situ hybridization to tissue sections, is the same in transformant and wild-type larvae. This indicates that all the genetic information specified by the w+ locus is contained within this 12-kb segment of DNA. By secondary mobilization it was shown that the w+ sequences have been inserted as a functional P(w+) transposon which is capable of further transposition.

Keywords: Drosophila, white gene, P-factor, transformation

Full text

PDF
2080

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BUTENANDT A., BICKERT E., KUEBLER H., LINZEN B. [On ommochromes. 20. On the distribution of ommatin in animals. New methods of identification and quantitative determination]. Hoppe Seylers Z Physiol Chem. 1960;319:238–256. doi: 10.1515/bchm2.1960.319.1.238. [DOI] [PubMed] [Google Scholar]
  2. Bender W., Spierer P., Hogness D. S. Chromosomal walking and jumping to isolate DNA from the Ace and rosy loci and the bithorax complex in Drosophila melanogaster. J Mol Biol. 1983 Jul 25;168(1):17–33. doi: 10.1016/s0022-2836(83)80320-9. [DOI] [PubMed] [Google Scholar]
  3. Bingham P. M., Levis R., Rubin G. M. Cloning of DNA sequences from the white locus of D. melanogaster by a novel and general method. Cell. 1981 Sep;25(3):693–704. doi: 10.1016/0092-8674(81)90176-8. [DOI] [PubMed] [Google Scholar]
  4. Clewell D. B., Helinski D. R. Properties of a supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry. 1970 Oct 27;9(22):4428–4440. doi: 10.1021/bi00824a026. [DOI] [PubMed] [Google Scholar]
  5. Engels W. R. Extrachromosomal control of mutability in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1979 Aug;76(8):4011–4015. doi: 10.1073/pnas.76.8.4011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Engels W. R. Germline hypermutability in Drosophila and its relation to hybrid dysgenesis and cytotype. Genetics. 1981 Jul;98(3):565–587. doi: 10.1093/genetics/98.3.565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gehring W. J., Paro R. Isolation of a hybrid plasmid with homologous sequences to a transposing element of Drosophila melanogaster. Cell. 1980 Apr;19(4):897–904. doi: 10.1016/0092-8674(80)90081-1. [DOI] [PubMed] [Google Scholar]
  8. Goldberg D. A., Posakony J. W., Maniatis T. Correct developmental expression of a cloned alcohol dehydrogenase gene transduced into the Drosophila germ line. Cell. 1983 Aug;34(1):59–73. doi: 10.1016/0092-8674(83)90136-8. [DOI] [PubMed] [Google Scholar]
  9. Goldberg M. L., Paro R., Gehring W. J. Molecular cloning of the white locus region of Drosophila melanogaster using a large transposable element. EMBO J. 1982;1(1):93–98. doi: 10.1002/j.1460-2075.1982.tb01130.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hafen E., Levine M., Garber R. L., Gehring W. J. An improved in situ hybridization method for the detection of cellular RNAs in Drosophila tissue sections and its application for localizing transcripts of the homeotic Antennapedia gene complex. EMBO J. 1983;2(4):617–623. doi: 10.1002/j.1460-2075.1983.tb01472.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hazelrigg T., Levis R., Rubin G. M. Transformation of white locus DNA in drosophila: dosage compensation, zeste interaction, and position effects. Cell. 1984 Feb;36(2):469–481. doi: 10.1016/0092-8674(84)90240-x. [DOI] [PubMed] [Google Scholar]
  12. JUDD B. H. Formation of duplication-deficiency products by asymmetrical exchange within a complex locus of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1961 Apr 15;47:545–550. doi: 10.1073/pnas.47.4.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jack J. W., Judd B. H. Allelic pairing and gene regulation: A model for the zeste-white interaction in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1368–1372. doi: 10.1073/pnas.76.3.1368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Langer-Safer P. R., Levine M., Ward D. C. Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4381–4385. doi: 10.1073/pnas.79.14.4381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Langer P. R., Waldrop A. A., Ward D. C. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. doi: 10.1073/pnas.78.11.6633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Levis R., Rubin G. M. The unstable wDZL mutation of Drosophila is caused by a 13 kilobase insertion that is imprecisely excised in phenotypic revertants. Cell. 1982 Sep;30(2):543–550. doi: 10.1016/0092-8674(82)90251-3. [DOI] [PubMed] [Google Scholar]
  17. O'hare K., Levis R., Rubin G. M. Transcription of the white locus in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6917–6921. doi: 10.1073/pnas.80.22.6917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pirrotta V., Bröckl C. Transcription of the Drosophila white locus and some of its mutants. EMBO J. 1984 Mar;3(3):563–568. doi: 10.1002/j.1460-2075.1984.tb01847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pirrotta V., Hadfield C., Pretorius G. H. Microdissection and cloning of the white locus and the 3B1-3C2 region of the Drosophila X chromosome. EMBO J. 1983;2(6):927–934. doi: 10.1002/j.1460-2075.1983.tb01523.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Richards G., Cassab A., Bourouis M., Jarry B., Dissous C. The normal developmental regulation of a cloned sgs3 'glue' gene chromosomally integrated in Drosophila melanogaster by P element transformation. EMBO J. 1983;2(12):2137–2142. doi: 10.1002/j.1460-2075.1983.tb01714.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rubin G. M., Spradling A. C. Genetic transformation of Drosophila with transposable element vectors. Science. 1982 Oct 22;218(4570):348–353. doi: 10.1126/science.6289436. [DOI] [PubMed] [Google Scholar]
  22. Scalenghe F., Turco E., Edström J. E., Pirrotta V., Melli M. Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes. Chromosoma. 1981;82(2):205–216. doi: 10.1007/BF00286105. [DOI] [PubMed] [Google Scholar]
  23. Scholnick S. B., Morgan B. A., Hirsh J. The cloned dopa decarboxylase gene is developmentally regulated when reintegrated into the Drosophila genome. Cell. 1983 Aug;34(1):37–45. doi: 10.1016/0092-8674(83)90134-4. [DOI] [PubMed] [Google Scholar]
  24. Spradling A. C., Rubin G. M. The effect of chromosomal position on the expression of the Drosophila xanthine dehydrogenase gene. Cell. 1983 Aug;34(1):47–57. doi: 10.1016/0092-8674(83)90135-6. [DOI] [PubMed] [Google Scholar]
  25. Spradling A. C., Rubin G. M. Transposition of cloned P elements into Drosophila germ line chromosomes. Science. 1982 Oct 22;218(4570):341–347. doi: 10.1126/science.6289435. [DOI] [PubMed] [Google Scholar]
  26. Van Deusen E. B. Sex determination in germ line chimeras of Drosophila melanogaster. J Embryol Exp Morphol. 1977 Feb;37(1):173–185. [PubMed] [Google Scholar]
  27. Wiley K., Forrest H. S. Terminal synthesis of xanthommatin in Drosophila melanogaster. IV. Enzymatic and nonenzymatic catalysis. Biochem Genet. 1981 Dec;19(11-12):1211–1221. doi: 10.1007/BF00484574. [DOI] [PubMed] [Google Scholar]
  28. Wilson T. G., Jacobson K. B. Isolation and characterization of pteridines from heads of Drosophila melanogaster by a modified thin-layer chromatography procedure. Biochem Genet. 1977 Apr;15(3-4):307–319. doi: 10.1007/BF00484462. [DOI] [PubMed] [Google Scholar]
  29. Zachar Z., Bingham P. M. Regulation of white locus expression: the structure of mutant alleles at the white locus of Drosophila melanogaster. Cell. 1982 Sep;30(2):529–541. doi: 10.1016/0092-8674(82)90250-1. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES