中文摘要：

目的 建立人醛糖还原酶（AR）及其与绿色荧光蛋白（GFP）融合表达的酿酒酵母细胞表达株，检测细胞中目的基因的表达及蛋白活性。方法将含目的基因AR及融合基因AR：：GFP的重组质粒pYEX—BX转化酵母细胞INVScl，得到宿主菌株XAR，XAG及对照菌株YEX。观察酵母细胞生长曲线及GFP荧光信号；Northemblot检测细胞中ARmRNA转录；Westernblot检测AR蛋白表达；荧光法测定AR活性。结果三菌株生长速率无明显差异；XAG菌株相对荧光强度与生长时间成线性关系；XAR，XAG菌株分别有持续、稳定的目的基因mRNA转录及目的蛋白表达，并具有AR活性。结论成功构建了AR基因的酵母表达株，为进一步将此细胞株应用于研究AR的致病机制及AR抑制剂的初步筛选打下基础。

英文摘要：

Objective To construct the Saccharomyces cerevisiae cell strains expressing aldose reductase（AR） and AR-green fluorescent protein（AR-GFP） fusion protein, and to detect the target gene expression and the AR enzyme activity. Methods The yeast expression plasmids pYEX-BX inserted with AR and AR：：GFP fusion gene were transformed into the yeast strain INVScl, which were named as XAR and XAG strains respectively. The blank pYEX-BX strain was used as the normal control. The growth curves and the fluorescence were determined in all strains. Northern blot, Western blot and the fluorescent method were used to detect the AR mRNA transcription, AR protein expression, and the AR activity respectively. Results There was no significant difference in the growth rates among three strains. There was a linear relationship between relative fluorescence of the XAG and the growth time. The mRNA transcription and protein expression of AR and AR：：GFP were sustainable and stable in XAR and XAG strains. The AR activities in the two strains were both proved by the fluorescent method. Conclusion The yeast expression strain of AR was constructed successfully, which lays basis for its application in the researches on pathogenic mechanism of AR and preliminary screening of new AR inhibitors.