中文摘要：

为了从工业生产菌株耐高渗产甘油假丝酵母（Candida gzycerinogenes）克隆甘油合成的限速酶编码基因胞浆NADT^＋3-磷酸甘油脱氢酶基因（ctGPD），对不同酵母和其他真核生物的NAD^＋-3-磷酸甘油脱氢酶进行比对，分析氨基酸和核苷酸的保守序列，设计了4对简并引物用于扩增C．gzycerinogenes的NAD^＋-3-磷酸甘油脱氢酶（GPD）基因片段，经过优化PCR反应条件，利用其中一对中等简并度的引物扩增出CgGPD基因中约600bp的保守核心片段。DNA序列及推绎的氨基酸序列进行比对分析表明，该基因片段与其他酵母的胞浆NADL3磷酸甘油脱氢酶基因的对应区域具有典型的保守区域，并且与安格斯毕赤酵母的GPD基因相似性较高。

英文摘要：

In Saccharomyces cerevisiae, glycerol is synthesized in cytosol by reduction of the glycolytic intermediate dihydroxyacetone phosphate （DHAP） to glycerol 3-phosphate （G3P） followed by dephosphorylation of G3P to glycerol. Further studies had revealed that cytol NAD^＋-dependent glycerol 3-phosphate dehydrogenase （ctGPD） is the rate-limiting enzymes for glycerol synthesis. However, it is unclear whether GPD has similar role in Candida glycerinogenes WL2002-5, a novel osmotolerant yeast species used in glycerol production. So, it is essential to clone GPD gene from C. glycerinogenes, for investigation of glycerol biosynthesis in molecular level. After comparison of the amino acid sequence of the GPDH proteins in several species, degenerate oligonucleoide primers were designed for PCR based on conservative regions in the amino acid sequence. PCR was performed on the genomic DNA of C. glvcerinogenes and parameters involved in amplification efficiency were optimized. Using one moderate degeneracy pair of primers, a degenerate PCR product of ca. O. 6 kb was amplified and sequenced. Which homologous to GPD1 of Saccharomyces cerevisiae. This provides a threshold of studying glycerol production pathway and osmotic stress response mechanism at genetic and molecular level in this yeast.