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. 1986 Apr;83(8):2579–2583. doi: 10.1073/pnas.83.8.2579

Molecular cloning of the muscle gene unc-22 in Caenorhabditis elegans by Tc1 transposon tagging.

D G Moerman, G M Benian, R H Waterston
PMCID: PMC323342  PMID: 3010313

Abstract

The previously described mutator system of Caenorhabditis elegans var. Bergerac has as one of its targets unc-22, a previously uncloned gene on chromosome IV important in assembly and function of the body wall musculature. By assuming that the mutator activity involved transposition of the repetitive element Tc1 into the unc-22 gene we have succeeded both in cloning the unc-22 gene and in demonstrating that Tc1 transposition is the principal basis of the mutator activity in the Bergerac strain. Although germ-line excision of Tc1 is sensitive to genetic background, somatic excision appears to be less so, suggesting that Tc1 movement is controlled differently in germ-line and somatic tissue. The availability of a transposon-based mutator system should aid in the cloning of additional genes in C. elegans, and the particular properties of this Tc1 system may provide information about the control of transposable element activity more generally.

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Selected References

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

  1. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dibb N. J., Brown D. M., Karn J., Moerman D. G., Bolten S. L., Waterston R. H. Sequence analysis of mutations that affect the synthesis, assembly and enzymatic activity of the unc-54 myosin heavy chain of Caenorhabditis elegans. J Mol Biol. 1985 Jun 25;183(4):543–551. doi: 10.1016/0022-2836(85)90170-6. [DOI] [PubMed] [Google Scholar]
  6. Eide D., Anderson P. Transposition of Tc1 in the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1756–1760. doi: 10.1073/pnas.82.6.1756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Emmons S. W., Klass M. R., Hirsh D. Analysis of the constancy of DNA sequences during development and evolution of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1333–1337. doi: 10.1073/pnas.76.3.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Emmons S. W., Yesner L. High-frequency excision of transposable element Tc 1 in the nematode Caenorhabditis elegans is limited to somatic cells. Cell. 1984 Mar;36(3):599–605. doi: 10.1016/0092-8674(84)90339-8. [DOI] [PubMed] [Google Scholar]
  9. Emmons S. W., Yesner L., Ruan K. S., Katzenberg D. Evidence for a transposon in Caenorhabditis elegans. Cell. 1983 Jan;32(1):55–65. doi: 10.1016/0092-8674(83)90496-8. [DOI] [PubMed] [Google Scholar]
  10. Epstein H. F., Waterston R. H., Brenner S. A mutant affecting the heavy chain of myosin in Caenorhabditis elegans. J Mol Biol. 1974 Dec 5;90(2):291–300. doi: 10.1016/0022-2836(74)90374-x. [DOI] [PubMed] [Google Scholar]
  11. Fedoroff N. V., Furtek D. B., Nelson O. E. Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Activator (Ac). Proc Natl Acad Sci U S A. 1984 Jun;81(12):3825–3829. doi: 10.1073/pnas.81.12.3825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ferguson E. L., Horvitz H. R. Identification and characterization of 22 genes that affect the vulval cell lineages of the nematode Caenorhabditis elegans. Genetics. 1985 May;110(1):17–72. doi: 10.1093/genetics/110.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goodman H. M., Olson M. V., Hall B. D. Nucleotide sequence of a mutant eukaryotic gene: the yeast tyrosine-inserting ochre suppressor SUP4-o. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5453–5457. doi: 10.1073/pnas.74.12.5453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Greenwald I. lin-12, a nematode homeotic gene, is homologous to a set of mammalian proteins that includes epidermal growth factor. Cell. 1985 Dec;43(3 Pt 2):583–590. doi: 10.1016/0092-8674(85)90230-2. [DOI] [PubMed] [Google Scholar]
  15. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Karn J., Brenner S., Barnett L., Cesareni G. Novel bacteriophage lambda cloning vector. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5172–5176. doi: 10.1073/pnas.77.9.5172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liao L. W., Rosenzweig B., Hirsh D. Analysis of a transposable element in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3585–3589. doi: 10.1073/pnas.80.12.3585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. MacLeod A. R., Waterston R. H., Brenner S. An internal deletion mutant of a myosin heavy chain in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5336–5340. doi: 10.1073/pnas.74.12.5336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. MacLeod A. R., Waterston R. H., Fishpool R. M., Brenner S. Identification of the structural gene for a myosin heavy-chain in Caenorhabditis elegans. J Mol Biol. 1977 Jul;114(1):133–140. doi: 10.1016/0022-2836(77)90287-x. [DOI] [PubMed] [Google Scholar]
  20. Mattox W. W., Davidson N. Isolation and characterization of the Beadex locus of Drosophila melanogaster: a putative cis-acting negative regulatory element for the heldup-a gene. Mol Cell Biol. 1984 Jul;4(7):1343–1353. doi: 10.1128/mcb.4.7.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Modolell J., Bender W., Meselson M. Drosophila melanogaster mutations suppressible by the suppressor of Hairy-wing are insertions of a 7.3-kilobase mobile element. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1678–1682. doi: 10.1073/pnas.80.6.1678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moerman D. G., Baillie D. L. Genetic Organization in CAENORHABDITIS ELEGANS: Fine-Structure Analysis of the unc-22 Gene. Genetics. 1979 Jan;91(1):95–103. doi: 10.1093/genetics/91.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moerman D. G., Plurad S., Waterston R. H., Baillie D. L. Mutations in the unc-54 myosin heavy chain gene of Caenorhabditis elegans that alter contractility but not muscle structure. Cell. 1982 Jul;29(3):773–781. doi: 10.1016/0092-8674(82)90439-1. [DOI] [PubMed] [Google Scholar]
  24. Moerman D. G., Waterston R. H. Spontaneous unstable unc-22 IV mutations in C. elegans var. Bergerac. Genetics. 1984 Dec;108(4):859–877. doi: 10.1093/genetics/108.4.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  26. Rogalski T. M., Baillie D. L. Genetic organization of the unc-22 IV gene and the adjacent region in Caenorhabditis elegans. Mol Gen Genet. 1985;201(3):409–414. doi: 10.1007/BF00331331. [DOI] [PubMed] [Google Scholar]
  27. Rogalski T. M., Moerman D. G., Baillie D. L. Essential genes and deficiencies in the unc-22 IV region of Caenorhabditis elegans. Genetics. 1982 Dec;102(4):725–736. doi: 10.1093/genetics/102.4.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rose A. M., Snutch T. P. Isolation of the closed circular form of the transposable element Tc1 in Caenorhabditis elegans. Nature. 1984 Oct 4;311(5985):485–486. doi: 10.1038/311485a0. [DOI] [PubMed] [Google Scholar]
  29. Rosenzweig B., Liao L. W., Hirsh D. Sequence of the C. elegans transposable element Tc1. Nucleic Acids Res. 1983 Jun 25;11(12):4201–4209. doi: 10.1093/nar/11.12.4201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ruan K., Emmons S. W. Extrachromosomal copies of transposon Tc1 in the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4018–4022. doi: 10.1073/pnas.81.13.4018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Searles L. L., Jokerst R. S., Bingham P. M., Voelker R. A., Greenleaf A. L. Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P element transposon tagging. Cell. 1982 Dec;31(3 Pt 2):585–592. doi: 10.1016/0092-8674(82)90314-2. [DOI] [PubMed] [Google Scholar]
  32. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  33. Sulston J. E., Brenner S. The DNA of Caenorhabditis elegans. Genetics. 1974 May;77(1):95–104. doi: 10.1093/genetics/77.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Waterston R. H., Thomson J. N., Brenner S. Mutants with altered muscle structure of Caenorhabditis elegans. Dev Biol. 1980 Jun 15;77(2):271–302. doi: 10.1016/0012-1606(80)90475-3. [DOI] [PubMed] [Google Scholar]
  35. Wills N., Gesteland R. F., Karn J., Barnett L., Bolten S., Waterston R. H. The genes sup-7 X and sup-5 III of C. elegans suppress amber nonsense mutations via altered transfer RNA. Cell. 1983 Jun;33(2):575–583. doi: 10.1016/0092-8674(83)90438-5. [DOI] [PubMed] [Google Scholar]

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