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以石蒜(Lycoris radiata)为试材,采用同源克隆和RACE的试验方法,克隆得到LrCMO基因全长cDNA,并对其进行了基因表达分析。结果表明:LrCMO基因全长1 472 bp,其中开放阅读框(ORF)为1 281 bp,编码427个氨基酸残基,预测编码蛋白质的分子量为47.92 kD,理论等电点为5.72;LrCMO是一个稳定的疏水蛋白,不具有跨膜结构,含有叶绿体导肽;LrCMO基因编码的氨基酸与植物其他CMO蛋白具有较高的一致性,且与海枣PdCMO、香蕉MaCMO及油棕EgCMO亲缘关系最高,聚为一类。实时荧光定量PCR分析表明,LrCMO在根、鳞茎和叶片中有表达,且在鳞茎中的表达量最高;LrCMO受聚乙二醇(PEG)处理的诱导表达,其基因相对表达量在处理后12 h达到最高。随着处理时间的延长,LrCMO基因相对表达量逐渐下调至对照水平。
Using Lycoris radiata as test material, the full-length cDNA of LrCMO gene was cloned by homologous cloning and RACE assay, and its gene expression analysis was carried out. The results showed that the LrCMO gene was 1 472 bp in length, including an open reading frame (ORF) of 1 281 bp encoding a protein of 427 amino acids with a predicted molecular mass of 47.92 kD and a theoretical isoelectric point of 5.72. LrCMO was a stable Of the hydrophobin did not have the transmembrane structure and contained the chloroplast leader peptide. The amino acids encoded by LrCMO gene were highly consistent with other plant CMO proteins and had the highest genetic relationship with PdCMO, banana MaCMO and oil palm EgCMO, one type. Real-time PCR analysis showed that LrCMO was expressed in roots, bulbs and leaves, and highest in the bulbs. LrCMO was induced by polyethylene glycol (PEG) treatment and its relative gene expression level was h reached the highest. With the extension of treatment time, the relative expression of LrCMO gene was gradually reduced to the control level.