油茶Pht1;1基因克隆及其表达分析

Isolation and Expression Analysis of CoPht1;1 from Oil Tea

以油茶湘林4号为材料,通过RT-PCR和RACE的方法克隆出油茶磷酸转运子Pht1基因家族一个成员的全长cD-NA序列,命名为CoPht1;1(GenBank登录号:JX403969),通过实时定量PCR的方法检测了不同磷浓度下该基因在根系中的表达水平。结果表明:CoPht1;1 CDS长度为1626 bp,编码542个氨基酸,与其他物种的Pht1氨基酸序列具有较高的相似性,其中与夹竹桃科长春花的Pht1相似性最高,达到88%;蛋白质二级结构和拓扑结构预测表明,CoPht1;1具有跨膜蛋白的主要特征,与其他物种的Pht1具有一致性;实时定量RT-PCR结果表明,油茶Pht1基因的表达受低磷诱导,并在受磷胁迫处理(P浓度为0.1 mmol/L)15 d时表达量最高。

The phosphate transporter named CoPhtl; 1 was isolated from oil tea（ Camellia oleifera Xianglin4） with the method of RT-PCR and RACE. Its expression under different phosphorus level at different time was checked with qRT-PCR. The CDS of CoPhtl; 1 is 1626 bp which codes a predicted protein of 542 amino acids. The amino acid identity compared with other phosphate transporters is highly conserved and showed the highest similarity （88%）with the Phtl; 1 of Catharanthus roseus. Putative secondary structure and topology of the encoding protein have the main feature of transmembrane protein which consistent with Phtl encoding protein in other species. The real-time RT-PCR result showed that expression of CoPhtl; 1 was induced by P-deficiency and reached a high level at 0.1 mmol/L phosphorus concentration after treated for 15 days.