Worm Breeder's Gazette 9(1): 21
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
Short genomic clones derived from the C. elegans in gene and a tropomyosin gene were obtained by screening an expression library with specific antibodies. The library was constructed by cloning short Sau3A fragments of genomic DNA into the plasmid vector pUR289, which has a polylinker site at the 3' end of the lacz gene. The identity of the tropomyosin clones was confirmed by sequencing of the cloned fragment. The fragment showed strong homology to rat skeletal alpha-tropomyosin and Drosophila muscle tropomyosin. The sequence of the paramyosin clone showed a hydrophobic amino acid repeat structure, consistent with the known alpha-helical coiled-coil structure of this protein To confirm the identity cf the putative paramyosin clone, Sau3A fragments from a bacteriophage lambda clone containing 14-20 kb of flanking sequence were cloned into lacz expression plasmid vector. Numerous new fragment were recovered which encoded paramyosin epitopes. One of these fragments reacted with a monoclonal antibody to paramyosin which also specifically reacted with a 24 kd CNBr-fragment. Comparison of the N-terminal sequence of the 24 kd CNBr-fragment and the deduced amino acid sequence of the fusion protein showed an exact match of 8 amino acids. Cosmid clones overlapping the paramyosin gene were also isolated. In RNA hybridization experiments, each probe detects a single RNA species of the expected size. The paramyosin mRNA is approximately the same size as the '28s' RNA from nematode Sequencing of the both genes is in progress in Japan.