Worm Breeder's Gazette 12(5): 77 (February 1, 1993)
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.
We have been working on how muscle gene effects on the phenotype of individual. Although total genome project is going on, molecular nature of muscle proteins are still obscure. We cloned cDNAs of troponin I and troponin C by screening cDNA library(Bob Barstead and Bob Waterston) with antitroponin I and antitroponin C against purified Ascalis proteins respectively. These antibodies crossreacted with the muscle structure and isolated proteins of C. elegans.
cDNA sequence of troponin I was the same as that of the both cDNA clones, cm15 e4 (96%; 290/302) and cm20 cl(93%;344/367). It was noted that MRC sequence was much fit to the final sequence than that of St Louis. Completed cDNA encoded 242 amino acids and had molecular mass of 27,558. In vertebrates, there are alternative splicing in the C-terminal half of the molecule. We have not yet known whether or not there is an isoform in the worm The final genome map located on YAC clone, Y52C9 ,the central of chromosome X. The genome sequencing is in progress by recloning fragment from cosmid clone.
Troponin C is one of calcium binding proteins in muscle. cDNA of the troponin C was cloned by expression cloning not by hybridization approaches with cDNA probe of chicken troponin C. Deduced amino acid was 161 amino acids. It is interested that the troponin C had three calcium binding domains. In vertebrate, there are four calcium bindings. Missing the first calcium binding domain in the worm might be some functional reasons. The genome mapping was performed by screening YAC filter with ECL(enhanced chemiluminescenst detection, Amersham) method and conventional radiolabeling(Alan Coulson). The results were consistent to each other. The final position of tnc-1 was close to the unc-38 (acetylcholine receptor) on the central of chromosome I. There could be a mutant of muscle related at region(Bob Waterston and Bem Wiliams). E-mail communication will accelerate to solve the question .