Studies on the ryanodine receptor gene, ryr-l, kra-l andunc-68 genes in the LG V Yasuji Sakube, Tatsuji Ikeda and Hiroaki Kagawa Molecular Biology, Department of Biology, Faculty of Science, Okayama University, Okayama Japan 700
c5191 8@jpnkudpc Ryr-l; The worm has the ryanodine receptor gene (vol. 12 #3 p.51, Vol. 13 #2
p58, Sakube etal.) and the functional protein could be the gene product (Vol. 12 #4 p.69 Maryon et a/.). We have completed the structure of the gene on the cosmid clone M04C11 that spans 30 kb and encodes 46 exons. The gene locates between CeLip-1 and odc-lLG V. To know the place of expressed gene, promoter-
lyr-1/lacZ fusion plasmids were injected into worms and were analyzed the activity of, beta-galactosidase. At the moment, the
ryr-1 gene was expressed in many tissues under the control of 5'UTS. Fusion plasmids having 4.8 k and 2.7 kb of 5'UTS were expressed in muscles. 1.5 kb fragment was expressed in intestine and nerve. Fragments having 1.3 kb and 0.8 kb were expressed in muscles or intestine and nerve cells. At the moment we have not a conclusive result. We also isolated cDNA clones from a library and products of RT PCR and constructed expression plasmids for antibody isolation. kra-l and
unc-68; Ando eta/. isolated a mutant kra-l (ketamine response abnormal) and mapped near
unc-68 LG V and speculated that a 0.5kb transcript might be a gene product of unc 68 (Vol. 12 #4 p.33 Ando et al.), but the result was over estimate. The recent experiment shows that mutation rescue with a gene fragment recloned from C46C8 was hard to tell the previous speculation. We also constructed a promoter/lacZ fusion plasmid for confirming the expression pattern of the gene fragment of the candidate and obtained this clone specifically expressed in the spermatocyte at L4 to young adult stages as was observed in Spe(sperm specific protein). We have to find another locus at the place of
unc-68. We are very much sorry about the previous result. Maryon et al. are isolating a deletion mutant of Iyr-1 by the method of Tc1 excision. We hope that analysis of different approach terminates the conclusion.