盐生植物小花碱茅外整流K^+通道PtSKOR基因的克隆及RNAi载体构建

Cloning Outward-Rectifying Potassium Channel PtSKOR Gene and Constructing Its RNAi Vector in Halophyte Puccinellia tenuiflora

为研究盐生植物小花碱茅(Puccinellia tenuiflora)耐盐的分子机制,本研究根据已克隆到的Pt SKOR基因片段,利用RACE技术得到小花碱茅Pt SKOR的3'和5'c DNA序列,拼接后的基因全长为2 353 bp,编码715个氨基酸,系统进化分析显示Pt SKOR编码外整流K+通道。在此基础上,采用酶切连接的方法,构建了Ca MV 35S启动子驱动的含Pt SKOR基因片段反向重复序列的Pt SKOR-RNAi植物表达载体。

In order to investigate the molecular mechanism of the halophyte plant Puccinellia tenuiflora for salt tolerance, the 3＆#39; cDNA sequence and 5＆#39; cDNA sequence were cloned with RACE technology based on the reported PtSKOR gene fragment. Sequencing results indicated that the length of the PtSKOR gene from P. tenuiflora was 2 353 bp encoding 715 amino acids. Homology comparison showed that it was the outward-rectifying potassium channel gene named PtSKOR. Then the RNAi plant expression vector PtSKOR-RNAi with inverted repeats of PtSKOR driven by the promoter CaMV 35S was constructed by restriction enzyme digestion and ligation.