巴西橡胶树胚胎晚期丰富蛋白基因的克隆及序列分析

Cloning and Sequence Analysis of the Late Embryogenesis Abundant Related Protein Gene from Hevea brasiliensis

根据抑制性消减杂交文库分离的EST片段,采用3'-RACE技术从巴西橡胶树胶乳中获得HbLEA14基因的全长cDNA序列,其ORF长456 bp,编码151个氨基酸,预测HbLEA14蛋白分子量为16.58 ku,等电点为5.10。同源性比对显示,HbLEA14与蓖麻RcLEA14、乳浆大戟EeLEA14、陆地棉GhLEA14、大豆GmLEA14、拟南芥AtLEA14、番茄SlER5的同源性分别为88%、80%、75%、73%、65%和61%,属于非典型第2组LEA蛋白。RT-PCR结果显示,死皮植株胶乳中HbLEA14的表达量明显高于健康树。

Using the primers designed from the known EST from a suppression subtractive hybridization-cDNA library,the full-length cDNA sequence of HbLEA14 was cloned from Hevea brasiliensis latex by 3’-RACE technique.The opening reading frame（ORF）of HbLEA14 was 456 bp in length,encoding a 151-amino-acid polypeptide with a predicted molecular weight of 16.58 ku and isoelectric point of 5.10.Cluster analysis revealed that HbLEA14 was an atypical LEA2,and showed 88% identity with Ricinus communis RcLEA14,80% to Euphorbia esula EeLEA14,75% to Gossypium hirsutum GhLEA14,73% to Glycine max GmLEA14,65% to Arabidopsis thaliana AtLEA14 and 61% to Solanum lycopersicum SlER5,respectively.In addition,HbLEA14 exhibited higher transcriptional expression level in the latex of tapping panel dryness trees than in healthy trees.This paper will benefit to further analyze the functions of HbLEA14.