A 38 kb segment containing thecdc2 gene from the left arm of fission yeast chromosome II: sequence analysis and characterization of the genomic DNA and cDNAs encoded on the segment

✍ Scribed by Machida, Masayuki; Yamazaki, Shuji; Kunihiro, Sumiko; Tanaka, Toshihiro; Kushida, Norihiro; Jinnno, Kouji; Haikawa, Yuji; Yamazaki, Jun; Yamamoto, Saori; Sekine, Mitsuo; Oguchi, Akio; Nagai, Yoshimi; Sakai, Mari; Aoki, Ken-ichi; Ogura, Keiko; Kudoh, Yutaka; Kikuchi, Hisashi; Zhang, Michael Q.; Yanagida, Mitsuhiro

• Publisher: John Wiley and Sons
• Year: 2000
• Tongue: English
• Weight: 104 KB
• Volume: 16
• Category: Article
• ISSN: 0749-503X
• DOI: 10.1002/(sici)1097-0061(20000115)16:1<71::aid-yea505>3.0.co;2-5

✦ Synopsis

A genomic 38 kbp segment on the c1750 cosmid clone containing the cdc2 gene, located in the left arm of chromosome II from Schizosaccharomyces pombe, was sequenced. The segment was found to have ®ve previously known genes, pht1, cdc2, his3, act1 and mei4. Among 11 coding sequences (CDSs) predicted by the gene ®nding software INTRON.PLOT., four CDSs, pi007, pi010, pi014 and pi016, had considerable similarity to 40S ribosomal protein, glycosyltransferase, cdc2-related protein kinase and a-1,2-mannosyltransferase, respectively. Another unusually huge open reading frame (ORF) (pi011), consisting of 2233 amino acids, existed, having signi®cant homology to a-amylase, granule-bound glycogen synthase and the Sz. pombe YS 1110 clone product at the N-terminal, middle and C-terminal regions, respectively. All the predicted 11 CDSs were experimentally analysed by RACE PCR. The sequencing of the RACE products revealed that there were two small overlaps at the 3k untranslated regions (UTRs) between pi004 and pi005 (17 bp) and between pi007 and pi008 (2 bp). The distances between 5k end of the 5kUTR and the putative translation initiation codon varied from 10 to 302 nucleotides (nt) among the nine CDSs successfully analysed by 5k-RACE. The expression level of each CDS on this clone was determined. Among the 16 genes on this clone, the previously determined genes, pht1, cdc2, his3 and act1, were found to be most highly expressed. Finally, cDNAs of all the newly identi®ed genes were detected by RACE, proving the actual expression of these genes.