Cloning and characterisation of therad9DNA repair gene fromSchizosaccharomyces pombe

Open Access
Publisher Website
Google Scholar
Abstract
The rad9. 192 DNA repair mutant from the fission yeast, Schizosaccharomyces pombe, is sensitive to both UV and lonising radiation. The rad9 gene has been cloned by complementatlon of the gamma-ray sensitivity of the mutant cell line. A 4.3kb Hindlll fragment was found to confer resistance to both types of radiation. The region of compiementation was further iocalised to a 2.6kb Hindlll-EcoRV fragment, which, by DNA sequence analysis, was found to contain sequences capable of coding for a 427 amino acid protein, if three introns were postulated to remove stop codons. The introns were confirmed by sequence analysis of cDNA clones and PCR products derived from cDNA. The product of transcription is a 1.6kb mRNA of low abundance. The putative rad9 protein shows no homology to any published sequence. A truncated protein is capable of complementing the radiation sensitivity of the rad9. 192 mutant. Deletion of the gene is not lethal and the null allele has a similar phenotype to the rad9. 192 mutant.