Proper expression of myosin genes in transgenic nematodes.
Caenorhabditis elegans has four genes which encode skeletal myosin heavy chain isoforms. We have re-introduced clones of two of these genes, myo-3 and unc-54 at low copy number into the germline of C. elegans. The resulting loci behave as functional copies of the genes by two genetic criteria: (i) they can result in phenotypic rescue of strains carrying inactivating myo-3 or unc-54 mutations, and (ii) their presence in strains with wild-type copies of the endogenous myosin loci has genetic consequences similar to duplicating the endogenous loci. The re-introduced genes function at a level close to that of the endogenous loci. Monoclonal antibodies specific for the different isoforms have been used to localize the expressed proteins. The re-introduced genes express in precisely the same cell types as the endogenous genes, and the myosin products produced assemble into filament structures as in wild-type. Unexpectedly, we have found in the course of this work that very high copy numbers of the unc-54 gene lead to a disruption of muscle structure which may result from overexpression of the protein product.
Fire
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Medical Research Council Laboratory of Molecular Biology, Cambridge, UK.
Waterston
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RH
D000818:Animals; D030801:Animals, Genetically Modified; D015139:Blotting, Southern; D002107:Caenorhabditis / Q000235:genetics*; D003433:Crosses, Genetic; D015870:Gene Expression; D005796:Genes; D005816:Genetic Complementation Test; D009154:Mutation; D009218:Myosins / Q000096:biosynthesis / Q000235:genetics*; D010641:Phenotype; D014162:Transfection; D014170:Transformation, Genetic
D009218:Myosins