doi:

DOI: 10.3724/SP.J.1264.2011.00014

Chinese Journal of Multiple Organ Diseases in the Elderly (中华老年多器官疾病杂志) 2011/10:4 PP.340-343

Induced expression and activity analysis of human aldose reductase gene in Saccharomyces cerevisiae


Abstract:
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 INVSc1, 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.

Key words:aldose reductase,yeast,green fluorescent protein,gene expression

ReleaseDate:2014-07-21 15:56:22



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