草菇Volvariella volvacea中乳清酸核苷-5’-单磷酸脱羧酶编码基因vv-pyrG的克隆和序列分析

Cloning and sequence analysis of the vv-pyrG gene encoding orotidine-5'monophosphate decarboxylase in Volvariella volvacea

运用简并PCR和基因组步行技术克隆常见食用菌草菇的乳清酸核苷-5’-单磷酸脱羧酶编码基因(vv-pyrG基因),并运用生物信息学和系统发育学的方法对所获得的基因进行了序列分析。结果显示:vv-pyrG基因的编码区长度为920 bp,含有2个长度分别为65 bp和54 bp的内含子,可以编码1个含有266个氨基酸残基的蛋白质序列。该氨基酸序列经比对分析,显示与裂褶菌(Schizophyllum commune)和灰盖鬼伞(Coprinus ci-nereus)的OMPDC序列的相同性分别为58%和64%,相似性分别为74%和78%。对24种真菌的OMPDC氨基酸序列进行系统发育分析表明,系统进化树中草菇与灰盖鬼伞、裂褶菌、双色蜡蘑(Laccaria bicolor)和白腐菌(Phanerochaete sordida)聚在一起,在节点上的支持率是97%。

A vv-pyrG gene （920 bp long） encoding orotidine-5 ＇-monophosphate decarboxylase （OMPDC）was cloned from Volvariella volvacea by means of nested degenerate PCR and genomic walking technology. The bioinformatic analysis indicated that the vv-pyrG gene had two introns being respectively 65 bp and 54 bp in length,and could encode a protein sequence containing 266 amino acid residues. The sequence alignment analysis of amino acids revealed that the identity and similarity be- tween the putative amino acid sequence encoded by the vv-pyrG gene and that of the OMPDC from Schizophyllum commune （or Coprinus cinereus） were 58% （or 64%） and 74% （or 78%）, respectively. The phylogenetic analysis on amino acid sequences of the OMPDC from 24 fungal species by the NJ method of the Phylip Software Package showed that Volvariella volvacea shared the same cluster with Laccaria bicolor ,S. commune, C. cinereus and Phanerochaete sordida ,and at the nodal point the supporting rate was 97% .