枸杞胆碱单加氧酶基因的克隆与表达分析

Molecular Cloning and Expression of CMO Encoding A Choline Monooxygenase in Lycium barbarum

根据GenBank中的植物胆碱加氧酶(CMO)基因的同源保守区设计简并引物,采用RT-PCR技术从枸杞中扩增出CMO基因的保守序列,并利用巢式PCR、5'-RACE和3'-RACE获得全长CMO。测序结果表明,CMO全长1540 bp,包含1 284 bp长的开放读码框(ORF),编码1个含427氨基酸残基、分子量为48.48 kDa、等电点(pI)为5.97的假定蛋白质。此序列Genbank登录号为FJ514800。mRNA分析表明,CMO基因在枸杞幼苗组织中受高盐胁迫和低温胁迫的上调诱导表达,表明该基因可能在植物抵御非生物胁迫中发挥重要作用。

Partial CMO cDNA was cloned from Lycium barbarum through RT-PCR technique by specific primers designed on the basis of CMO gene on GenBank website. Moreover, a 1 540 bp full length cDNA sequence of CMO was obtained using nested PCR, 5′- RACE and 3′-RACE technique. Further analyses on CMO indicated that an open reading frame （ORF） of 1 284 bp contained in this gene and predicted to encode a protein of 48.48 kDa with 427 amino acids. The gene accession nucleotide sequence number in GenBank was FJ514800. Semi-quantitative RT-PCR assay revealed that CMO was significantly up-regulated in Lycium barbarum under salt and cold stresses. It suggests that CMO might play an important role in Lycium barbarum＇s responses to abiotie stresses.