GENOME ANALYSIS OF CHROMOSOME III OF FISSION YEAST

• Published in: Genes & Genetic Systems Vol. 72; no. 6; p. 407
• Main Authors: HONGO Etsuko, MORIMYO Mitsuoki, MITA Kazuei, AJIMURA Masahiro, HIGASHI Tomoyasu, SUGAYA Kimihiko, SASANUMA Shunichi, NOHATA Junko, KIMURA Terumi, INOUE Hiromi, ISHIHARA Yoshie, KOIKE Shizuko
• Format: Journal Article
• Language: Japanese
• Published: The Genetics Society of Japan 1997
• ISSN: 1341-7568; 1880-5779

In a study of structures and functions of housekeeping genes of eukaryotic cells, we have chosen S. pombe as a model organism and made a catalog of 2,500 S. pombe cDNAs by sequencing 12,000 cDNA clones made from mRNA. If they are evenly scattered on the chromosome, 600 cDNA clones will be mapped on the chromosomeIII because it is one-fourth of the total genome size and the average 6 kb walking is enough to determine its complete DNA sequence. By using 200 cDNA and 200 genomic clones mapped on chromosome III, we have been sequencing the chromosome III of S. pombe by a bottom-up strategy of "walking" with using them as anchor sites. We have sequenced about 40 lambda clones carrying 15 kb DNA of S. pombe chromosome III and identified 120 ORFs as a gene from homology search in protein databases and in cDNA clones obtained by us. Among them, 15 genes related to radiation sensitivity were tried to disrupt them by transposon Tf1 insertion and the gene disruption was confirmed by PCR using one primer in Tf1 transposon and another primer flanking to the relevant gene.