筇竹CtHAK1基因的克隆及表达分析

Cloning and Expression Analysis on CtHAK1 Gene from Chimonobambusa tumidissinoda

为了研究钾转运载体HAK家族基因在竹笋以及竹子运输钾的特征,以筇竹实生苗(2 a)为研究材料,克隆出钾转运载体基因,命名为CtHAK1(基因登录号为:MT078978),并分析其表达特征。结果表明,CtHAK1基因全长为2633 bp,开放阅读框为2361 bp,编码786个氨基酸,相对分子量为87.91 kD,理论等电点(PI)为8.82。聚类分析显示CtHAK1与芦苇同源性最高,达到93.88%,亚细胞定位预测主要存在于细胞质膜上(60%)。RT-PCR结果显示,CtHAK1基因在不同组织(笋、根、茎、叶)中均有表达为非诱导表达,表达量由大到小为茎、根、叶、笋。KCl处理下筇竹根、茎部基因表达呈下调趋势,而叶呈上调表达,并且处理过程叶部的表达量始终高于根部和茎部的表达量。

In order to study the characteristics of potassium transporter HAK family gene in bamboo shoots and bamboos,a potassium carrier gene named CtHAK1(named as MT078978)was cloned from a 2-year seedling of Chimonobambusa tumidissinoda and its expression characteristics were analyzed.The results showed that the full length of CtHAK1 gene was 2633 bp,and the open reading frame(ORF)of CtHAK1 was 2361 bp,encoding 786 amino acids.The relative molecular weight of CtHAK1 protein is 87.91 kD,with the theoretical isoelectric point 8.82.Cluster analysis showed that CtHAK1 has the highest homology with Phragmites australis.In addition,subcellular localization predicted that this protein mainly located in cytoplasmic membrane(60%).The results by RT-PCR showed that CtHAK1 gene was expressed in different tissues(shoots,roots,stems and leaves),which is a non-induced form.And the expression amount of CtHAK1 was stem>root>leaf>shoot.The CtHAK1 of expression level in roots and stems was down-regulated,while in leaves was up-regulated under KCl treatment,and that in leaves was always higher than that in roots and stems.