甘蔗单糖转运蛋白基因ShPST2a克隆及亚细胞定位

Cloning and Subcellular Localization Analysis of Monosaccharide Transporter in Sugarcane

参照GenBank中公布的甘蔗品种Q117ShPST2a基因序列,设计特异性引物,采用RT-PCR方法从甘蔗成熟茎秆中扩增出甘蔗单糖转运蛋白的基因序列,命名为ShPST2a,该序列长2 455 bp,包含1个2 238 bp阅读框,编码745个氨基酸;构建该基因的瞬时表达载体,通过基因枪轰击洋葱表皮的方法对ShPST2a编码蛋白进行亚细胞定位研究。结果表明,该基因编码蛋白定位于细胞膜,符合细胞膜转运蛋白的特征,为进一步研究该基因编码蛋白的结构和功能奠定了基础。

ShPST2a gene was amplified by RT-PCR from mature stalks of sugarcane based on ShPST2a gene sequence of sugarcane variety Ql17 in Genbank. Subcellular localization was then used to study the function of ShPST2a. The plant expression vector pCAMBIA-ShPST2a with green fluorescent proteins （GFP） was constructed and transformed into onion epidermal cells via gene gun method. GFP fluorescence observation indicated that the ShPST2a-GFP fusion protein was located in the plasma membrane, thus ShPST2a encoding protein is a kind of membrane protein.