【摘要】 根据已获得的盐生植物獐茅(Aeluropus littoralisvar.sinensisDebeaux)液泡膜Na+/H+逆向转运蛋白(Na+/H+antiporter)基因部分cDNA片段,设计2条特异引物(GSP1、GSP2),采用RLM-RACE(RNA ligase-medi-ated rapid amplification of 5′and 3′cDNA ends)法获得了其5-′cDNA末端序列,并找出了转录起始位点。该片段长816 bp,与芦苇液泡膜Na+/H+逆向转运蛋白基因序列同源性最高达91%,与拟南芥、盐角草、水稻、大麦、小麦的同源性分别为81%、82%、86%、87%和81%,为该基因全长的克隆及启动子的研究奠定了基础。与其它测定基因转录起始位点的方法相比,RLM-RACE方法更快捷、简便、准确。更多还原

【Abstract】 RLM-RACE is a new technique of amplifying gene ends.Based on part of known cDNA sequence,two specific PCR primers were designed.The 5′-cDNA ends of Na+/H+ antiporter gene on the tonoplast of Aeluropus littoralis were cloned by RLM-RACE and the transcription initiation site was detected.The amplified fragment was 816bp in length and the nucleotide sequence analysis showed 91% similarity to the tonoplast Na+/H+ antiporter gene of Phragmites australis(PcNHX1),81% to Arabidopsis thaliana(AtNHX1),82% to Salicornia europaea(SeNHX1),86% to Oryza sativa(OsNHX1),87% to Hordeum vulgare(HvNHX1)and 81% to Triticum aestivum(TaNHX1).The result provided foundation for cloning the full sequence and studying the promoter of the gene.RLM-RACE(RNA ligase-mediated rapid amplification of 5′ and 3′ cDNA ends) is more rapid and simple comparing with the common methods for determining gene transcription initiation sites.更多还原