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目的:克隆和分析油茶高亲和磷转运蛋白基因,为研究其结构和功能打下基础。方法:以油茶品种‘华硕’为试材,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;2(GenBank登录号:JX412956.1),通过生物信息学技术对其序列的理化性质、结构与功能进行分析和预测。结果:CoPht1;2 CDS长度为1 590bp,编码530个氨基酸,与其它物种的Pht1氨基酸序列具有较高的相似性,其中与杨柳科毛果杨的Pht1相似性最高,达到77.5%;该基因所编码蛋白质的分子量为58.02 kDa,理论等电点pI为8.97,二级结构主要由α-螺旋、β-折叠和不规则卷曲构成,包含12个明显的跨膜螺旋拓扑结构。结论:预测显示该蛋白是一个疏水跨膜蛋白,具有磷转运蛋白的主要特征,初步判定其与油茶磷吸收有关,其功能有待进一步验证。
Objective: To clone and analyze the gene of Camellia oleifera high affinity and phosphate transporter, and lay the foundation for the study of its structure and function. Methods: The full-length cDNA sequence of Pht1 gene family was cloned by RT-PCR and RACE from Camellia oleifera ’ASUS’, named CoPht1; 2 (GenBank accession number: JX412956.1 ), Through the bioinformatics technology of its sequence of physical and chemical properties, structure and function analysis and prediction. Results: The CDS of CoPht1; 2 CDS was 530 bp in length and encoded a polypeptide of 530 amino acids with high similarity to Pht1 amino acid sequences of other species. The deduced amino acid sequence of CoPht1; 2 had the highest similarity with that of Populus tomentosa, reaching 77.5% The molecular weight of the encoded protein was 58.02 kDa and its theoretical isoelectric point was 8.97. The secondary structure consisted mainly of α-helix, β-sheet and irregular curl, and contained 12 obvious transmembrane helical topologies. CONCLUSION: It is predicted that this protein is a hydrophobic transmembrane protein with the main features of phosphorus transporter. It is preliminarily determined that it is related to phosphorus absorption of Camellia oleifera, and its function needs to be further verified.