国家科技攻关计划
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cDNA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 1 626 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基因的表达受低磷因素诱导,并随磷处理时间的不同而变化。
The phosphate transporter named CoPht1;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 CoPht1; 1 is 1 626 bp in length and it codes a predicted protein of 542 amino acids. The amino acid identity compared with other phosphate transporters is highly conserved and shows the highest similarity(88%)with the CoPht1;1 of Catharanthus roseus. Putative secondary structure and topology of the encoding protein have the main feature of transmembrane protein, and is consistent with Pht1 encoding protein in other species. The real-time RT-PCR result showed that expression of CoPht1;1 is induced by P-deficiency,and varied at different time. The results showed that CoPht1; 1 play an important role in phosphate uptake in camellia.
袁军,周俊琴,谭晓风.油茶Pht1;1基因克隆及其表达分析[J].植物遗传资源学报,2013,14(3):512-517.
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