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Molecular cloning and characterization of the dpy-20 gene of Caenorhabditis elegans

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Abstract

We describe the molecular analysis of the dpy20 gene in Caenorhabditis elegans. Isolation of genomic sequences was facilitated by the availability of a mutation that resulted from insertion of a Tc1 transposable element into the dpy-20 gene. The Tc1 insertion site in the m474:: Tc1 allele was identified and was found to lie within the coding region of dpy-20. Three revertants (two wild-type and one partial revertant) resulted from the excision of this Tc1 element. Genomic dpy-20 clones were isolated from a library of wild-type DNA and were found to lie just to the left of the unc-22 locus on the physical map, compatible with the position of dpy-20 on the genetic map. Cosmid DNA containing the dpy-20 gene was successfully used to rescue the mutant phenotype of animals homozygous for another dpy-20 allele, e1282ts. Sequence analysis of the putative dpy-20 homologue in Caenorhabditis briggsae was performed to confirm identification of the coding regions of the C. elegans gene and to identify conserved regulatory regions. Sequence analysis of dpy-20 revealed that it was not similar to other genes encoding known cuticle components such as collagen or cuticulin. The dpy-20 gene product, therefore, identifies a previously unknown type of protein that may be directly or indirectly involved in cuticle function. Northern blot analysis showed that dpy-20 is expressed predominantly in the second larval stage and that the mRNA is not at all abundant. Data from temperature shift studies using the temperature-sensitive allele e1282ts showed that the sensitive period also occurs at approximately the second larval stage. Therefore, expression of dpy-20 mRNA and function of the DPY-20 protein are closely linked temporally.

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Author notes

  1. Denise V. Clark

    Present address: Department of Biology, University of New Brunswick, (Bag Service 45111), E3B 6E1, Fredericton, N.B., Canada

Authors and Affiliations

  1. Institute of Molecular Biology and Biochemistry, Department of Biological Sciences, Simon Fraser University, V5A 1S6, Burnaby, B.C., Canada

    Denise V. Clark, Dinar S. Suleman, Karen A. Beckenbach, Erin J. Gilchrist & David L. Baillie

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  1. Denise V. Clark
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  2. Dinar S. Suleman
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  3. Karen A. Beckenbach
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  4. Erin J. Gilchrist
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  5. David L. Baillie
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Communicated by D. J. Finnegan

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Clark, D.V., Suleman, D.S., Beckenbach, K.A. et al. Molecular cloning and characterization of the dpy-20 gene of Caenorhabditis elegans . Molec. Gen. Genet. 247, 367–378 (1995). https://doi.org/10.1007/BF00293205

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  • DOI: https://doi.org/10.1007/BF00293205

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