巴西橡胶树HbHMAD1的克隆及表达

Cloning and Expression of HbHMAD1 from Hevea brasiliensis

根据已获得的在橡胶树自根幼态无性系与老态无性系胶乳中一个差异表达的片段序列设计引物,通过RACE法获得了橡胶树编码含有重金属相关结构域蛋白的cDNA,命名为HbHMAD1。序列分析结果表明,HbHMAD1长为814 bp,含有453 bp的阅读框,78 bp的5'-UTR和283 bp的3'-UTR,编码150个氨基酸,分子量为16.9 ku,等电点为10.4。HbHMAD1含有重金属相关结构域,与蓖麻(Ricinus communis)、拟南芥(Arabodopsis thesis)(3个)和玉米(Zea mays)中的含有重金属相关结构域蛋白的同源性分别为85%、64%、63%、60%和55%。半定量RT-PCR分析表明HbHMAD1基因在胶乳和树皮中表达,在叶中微量表达,在花中基本不表达。

The eDNA coding heavy-metal-associated domain protein, designated as HbHMAD1, was isolated from Hevea brasiliensis by rapid amplification of eDNA ends. HbHMAD1 consisted of a 453 bp open reading frame encoding 150 amino acids with molecular weight of 16.9 ku, a 78 bp 5＂UTR and a 283 bp 3＂UTR,. The deduced amino acid sequence of HbHMADi showed high identity of 85%, 64%, 63%, 60% and 55% to those of the heavy-metal-associated domain protein from Ricinus communis, Arabidopsis thaliana and Zea mays, respectively. Semi-quantitative reverse transcription-polymerase chain reaction analysis revealed that HbHMAD1 was expressed more in the latex than in the barks, whereas little expression was detected in leaves and flower.