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SeNHX1表达蛋白的亚细胞定位及其对Na~+的解毒功能

DOI: 10.3969/j.issn.1000-7091.2012.05.002, PP. 8-11

Keywords: SeNHX1基因,酵母,表达载体,GFP绿色荧光,耐盐性,亚细胞定位

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Abstract:

将带有目的基因SeNHX1、GFP的酵母表达载体pYES2-SeNHX1、pYES2-GFP、pYES2-SeNHX1-GFP分别导入nhx1酵母缺失型菌株296H,用共聚焦荧光显微镜观察SeNHX1表达蛋白的亚细胞定位;通过菌株在含盐培养基中生长状况研究该蛋白对Na+的解毒功能。结果表明,296H(pYES2-GFP)菌株的绿色荧光弥散于整个细胞质,而296H(pYES2-SeNHX1-GFP)菌株的绿色荧光包围于液泡膜的一圈,因此,SeNHX1蛋白定位于液泡膜;296H(pYES2-GFP)和296H缺陷型菌株在含0.5mol/LNaCl的培养基中生长受到抑制;带有SeNHX1目的基因的转化子比以上2种菌株在含盐培养基中生长更快,表明SeNHX1基因可恢复nhx1酵母突变体Na+的解毒功能。SeNHX1疏水性分析、氨基酸序列分析、亚细胞定位及其对Na+的解毒功能结果说明,SeNHX1基因是可提高生物体耐盐性的液泡膜Na+/H+逆向转运蛋白基因。

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