长穗偃麦草HKT1;4基因片段的克隆及序列分析

Cloning and Sequence Analysis of the HKT1;4 Gene Fragment in Elytrigia elongata

根据已报道的其他植物高亲和钾离子转运蛋白(HKT1;4)基因的保守序列设计一对简并性引物,以长穗偃麦草幼根总RNA为模板,采用RT-PCR方法克隆出长穗偃麦草HKT1;4,基因命名为EeHKT1;4。结果表明:该基因片段长度为614 bp,编码204个氨基酸,所得序列与其他植物氨基酸序列同源性均在80%以上,特别是和一粒小麦TmHKT1;4氨基酸同源性高达92%。这为长穗偃麦草HKT1;4全长基因的克隆及其耐盐分子机制的研究奠定基础。

Degenerate primers were designed based on the conserved sequences of the high-affinity K＋transporter（HKT1;4） genes from other plants. Total RNA was extracted from the roots of Elytrigia elongata to obtain an HKT1;4gene fragment by reverse transcription polymerase chain reaction（RT-PCR）, which named it as EeHKT1;4. Results showed that the length of EeHKT1;4 gene fragment was 614 bp encoding a 204 amino acids. The deduced amino acid sequence of EeHKT1;4 shared over 80% amino acid homology compared with the HKT1;4 from other plants, and especially shared over 92% amino acid homology compared with the TmHKT1;4 of Triticum monococcum. This study would lay the foundation for cloning full-length HKT1;4 gene and molecular mechanism of salt-tolerance in E. elongata.