A 38 kb segment containing the cdc2 gene from the left arm of fission yeast chromosome II: sequence analysis and characterization of the genomic DNA and cDNAs encoded on the segment

A genomic 38 kbp segment on the c1750 cosmid clone containing the cdc2 gene , located in the left ann of chromosome II from Schizosaccharomyces pombe, was sequenced, The segment was found to have five previously known genes, p ht1, cdc2, his3, act1 and mei4. Among 11 coding sequences (CDSs) predicted by the gene finding software INTRON.PLOT., four CDSs, pi007, pi010, pi014 a nd pi016, had considerable similarity to 40S ribosomal protein, glycosyltra nsferase, cdc2-related protein kinase and alpha-1,2-mannosyltransferase, re spectively. Another unusually huge open reading frame (ORF) (pi011), consis ting of 2233 amino acids, existed, having significant homology to a-amylase , granule-bound glycogen synthase and the Sz. pombe YS 1110 clone product a t the N-terminal, middle and C-terminal regions, respectively. All the pred icted 11 CDSs were experimentally analysed by RACE PCR. The sequencing of t he RACE products revealed that there were two small overlaps at the 3' untr anslated regions (UTRs) between pi004 and pi005 (17 bp) and between pi007 a nd pi008 (2 bp). The distances between 5' end of the 5'UTR and the putative translation initiation codon varied from 10 to 302 nucleotides (nt) among the nine CDSs successfully analysed by 5'-RACE. The expression level of eac h CDS on this clone was determined. Among the 16 genes on this clone, the p reviously determined genes, pht1, cdc2, his3 and act1, were found to be mos t highly expressed. Finally, cDNAs of all the newly identified genes were d etected by RACE, proving the actual expression of these genes. The nucleoti de sequence has been submitted to the EMBL database under Accession No. AB0 04534. Copyright (C) 2000 John Wiley & Sons, Ltd.