Expression and Detection the Enzyme Activity of the Wild and Mutation Type of CYP51 Protein of Candida albicans / 微生物学通报

The Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R mutants of Candida albicans sterol 14?-demethylase (CACYP51) were constructed and heterologously expressed in D12667, the reconstructed strain with the deletion of CYP51 gene of the Y12667. With the strains obtained and microsome enzymes separated, the western blot and the ultraviolet absorption spectrophotometry were used to qualitative and quantitative detect the expressed protein, the GC-MS was used to detect the metabolism activity of the protein. The results showed that, the target protein expressed successfully in the reconstructed strains, with the expression level up to 25% of the total microsome proteins. The results also showed that, the wild type protein had the catalytic activity to its nature substrate. While after alteration the wild gene with Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R by a single base substitution, the catalytic activity of protein markedly decreased respectively. So the wild type and mutation CYP51 were expressed successfully in Saccharomyces cerevisiae and the expression products preserved the activity to metabolism their nature substrate.

THE EFFECTS OF HEPATOCYTE GROWTH FACTOR AND VASCULAR ENDOTHELIAL GROWTH FACTOR ON PROLIFERATION OF VASCULAR WALL CELLS / 解放军医学杂志

The effects of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) on proliferation of bovine coronary artery endothelial cells (BCAEC) and smooth muscle cells (BCASMC) were studied in vitro. BCAEC and BCASMC were isolated and cultured and divided into control group, VEGF (50ng/ml) group and HGF (50ng/ml) group. Cells proliferation was measured using MTT method. The results showed that the OD values of control, VEGF, and HGF group in BCAEC cultures were 0.23?0.02, 0.58?0.10, and 0.42?0.12, respectively, and those in BCASMC were 0.31?0.08, 0.45?0.09, and 0.40?0.11, respectively. The proliferation ratios of BCAEC and BCASMC induced by HGF were 152.2%?33.8% and 45.2%?25.3%, respectively, and that by VEGF were 82.6%?18.7% and 29.0%?20.4%, respectively. The results suggested that HGF could promote proliferation and migration of BCAEC and BCASMC, while VEGF could promote proliferation of BCAEC but not BCASMC. The effect of HGF on BCAEC was stronger than that on BCASMC, and the induction strength of HGF was higher than VEGF.