橙色红曲菌乳清苷-5'-磷酸脱羧酶基因的克隆

Cloning of Orotidine-5′-phosphate decarboxylase gene from Monascus aurantiacus

根据已报道的真菌乳清苷-5′-磷酸脱羧酶(pyrG)氨基酸序列,采用CODEHOP策略设计简并引物,从橙色红曲菌(Monascus aurantiacus)的基因组DNA中克隆得到pyrG基因片段。然后运用PCR法筛选橙色红曲菌Fosmid文库,获得了pyrG基因全长(GenBank登录号:GU723506)。经过blastx比较发现,橙色红曲菌pyrG基因与AspergillusflavusNRRL3357的氨基酸序列同源性最高,达到81%。该基因能够作为筛选标记应用于橙色红曲菌同源转化系统。

According to the known Orotidine-5′- phosphate decarboxylase, pyrG gene fragment was cloned from Monascus aurantiacus genomic DNA using degenerate primers designed with CODEHOP strategy. And its complete sequence was obtained by a PCR-based method for screening fosmid library（Accession No.GU723506）. After the blastx comparison,the cloned gene was confirmed that it was Monascus pyrG gene with 81% sequence identity to that of Aspergillus flavus NRRL3357. It can use as a selective marker for Monascus aurantiacus homologus transformation system.