高渗工业菌株Candida glycerinogenes整合型表达载体的构建及验证

Development and Verification of an Integrative Vector for the Industrial Osmotolerant Yeast Candida glycerinogenes

DOI：10.3969/j.issn.1673-1689.2015.08.009

中文关键词: 产甘油假丝酵母 3-磷酸甘油脱氢酶基因启动子 绿色荧光蛋白 整合型载体 高渗表达

英文关键词: Candida glycerinogenes CGMCC NO. 6830, glycerol-3-phosphate dehydrogenase promoter, green fluorescent protein, integrative vector, high osmotic pressure expression

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中文摘要:

构建应用于工业用产甘油假丝酵母（Candida glycerinogenes） CGMCC NO. 6830的整合型表达载体，建立一种可利用底物形成的渗透压调控表达外源基因的体系。以pUC19质粒为基本骨架，C. glycerinogenes CGMCC NO. 6830的18S rDNA为整合位点，运用其3-磷酸甘油脱氢酶基因启动子PCggpd启动外源基因表达，腐草霉素抗性基因ble作为重组酵母转化子筛选标记，获得应用于产甘油假丝酵母的稳定的表达型整合载体；以绿色荧光蛋白基因gfp作为报告基因考察该表达质粒的性能，实现了外源基因gfp的渗透压调控表达。

英文摘要:

To express heterogeneous genes under the regulation of osmotic pressure in Candida glycerinogenes CGMCC NO. 6830，an integrative vector was constructed in this study. The constructed integrative vector harbored the backbone of plasmid pUC19，18S rDNA sequence of C. glycerinogenes CGMCC NO. 6830 as the integration site，phleomycin-resistant gene（ble） as a selectable marker，the osmolarity regulated promoter（PCggpd） and green fluorescent protein gene gfp as a reporter. The target gene was integrated into chromosomes of C. glycerinogenes CGMCC NO. 6830 and the expression level of gfp was regulated by osmotic pressure.

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