新型渗透压调控的工业酵母启动子

New osmo-regulational promoters in the industrial yeast

摘要:

摘要:【目的】筛选工业酵母Candida glycerinogenes渗透压调控性能优越的启动子，为工业酵母改造及转基因研究提供新途径。【方法】PCR扩增C. glycerinogenes新型系列启动子PCgPGI、PCgTPI、PCgZWF、PCgSTL1、PCgSTL2、PCgSTL3，利用生物信息学技术解析启动子序列中渗透压胁迫应答元件，构建包含gfp荧光蛋白报告基因和PCgPGI、PCgTPI、PCgZWF、PCgSTL1、PCgSTL2、PCgSTL3启动子的5.8S rDNA整合表达载体，通过荧光强度及mRNA转录的qRT-PCR测定结果检验各启动子活性强度及其受渗透压调控情况。【结果】启动子PCgSTL3包含多个STRE渗透压胁迫应答元件，在工业酵母中受渗透压调控更敏感，启动强烈，转录水平高，gfp表达量大。【结论】PCgSTL3是受渗透压调控能够实现外源基因可控表达的优良工业酵母启动子。

Abstract:

Abstract:［Objective］The work was aimed at selecting osmo-regulated prompters possessing excellent performance for further research of the industrial yeast Candida glycerinogenes.［Methods］Promoters PCgPGI，PCgTPI，PCgZWF，PCgSTL1，PCgSTL2 and PCgSTL3 were amplified by PCR and their bioinformatics analysis of stress response elements (STREs) were conducted． We constructed integrative plasmids containing 5. 8S rDNA，a fluorescence protein gene gfp and a promoter PCgPGI，PCgTPI，PCgZWF，PCgSTL1，PCgSTL2 or PCgSTL3．The promoters’activities and osmoregulations were compared according to the results of fluorescence and qRT-PCR.［Results］PCgSTL3 had more STREs，higher transcription level，lager gfp expression and it was more sensitive to stress.［Conclusion］PCgSTL3 is an excellent induced promoter responding to hyperosmotic stress． Controlled expression of target genes can be realized using PCgSTL3 in the industrial yeast.