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.

Exson-expression cloning of the unc-15 paramyosin gene and a tropomyosin gene from Caenorhabditis elegans

H. Kagawa, J. Karn, and S. Brenner

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 
Sequencing of the both genes is in progress in Japan.