Cloning Of PyrG Gene Encoding Orotidine-5'-monophosphate Decarboxylase In Blakeslea Trispora Used Genome Walker And Sequence Analyse

Based on the homology analysis of nucleic acid sequence encoding Orotidine-5'-monophosphate Decarboxylase from Mucor circinelloides,,Phycomyces blakesleeanus,Rhizopus niveus and Rhizomucor pusillus, in the third exon, a pair of primers was designed. Used the primers, an approximate 500bp product was obtained from Blakeslea trispora genome by PCR. Through homology analysis, it is proved that the product is the gene fragment encoding Orotidine-5'-monphosphate Decarboxylase. Then walking upstream and downstream in genomic DNA from the known sequence was proceeded by a suppression PCR . The flanking DNA fragments were amplified and cloned by PCR respectively, and the whole gene was sequenced completely. The result indicated that homology of the amino acid sequence in exons, compared with the Mucor circinelloides, Phycomyces blakesleeanus and Rhizomucor pusillus, were 89.8%, 86.4ï¼… and 86.5ï¼…. The gene could be used to construct the transformation system of Blakeslea trispora.