红海榄脱水素基因(RsDHN1)的克隆及表达分析

采用RT-PCR和RACE技术对已获得的红海榄根部其中一个差异表达基因进行克隆,通过Blast分析和序列比对可知,该基因全长cDNA序列包括146bp的5'端非编码区,43bp的3'端非编码区和一个编码241个氨基酸的开放阅读框,命名为RsDHN1基因。该cDNA编码的蛋白具有植物脱水素蛋白的特征性结构,属于SKn类脱水素蛋白,与其它植物的SKn类脱水素蛋白具有43%～55%的氨基酸序列同源性。半定量RT-PCR分析表明,盐胁迫对RsDHN1基因的表达有明显上调作用。

Cloning and Characterization of Dehydrin Gene of Rhizophora stylosa(RsDHN1)

Rhizophora stylosa is one of the most common species in Hainan mangrove forest. In order to identify salt-tolerant-related genes form Rhizophora stylosa, seedlings of Rhizophora stylosa were planted in fresh water or saline. The differentially expressed ESTs(expressed sequence tag) were detected by macroarry in roots of Rhizophora stylosa. Full-length sequence of the EST was identified by RACE techniques(rapid amplification of cDNA ends). It had 943 bp, which contained a 146 bp 5'-UTR(untranslation region), a 43-bp 3'-UTR and an open reading frame of 726 bp, which encoded a protein of 241 amino acid residues. The deduced protein showed a typical domain of plant dehydrin proteins and contains the conserved S and K segments that belonged to SKn class of plant dehydrin. The gene was termed as RsDHN1. RsDHN1 shared 43%~55% identity with the SKn dehydrins of other plant species. RT-PCR analysis showed that salt treatment significantly upregulated the expression of RsDHN1.