摘要
将酿酒酵母的PSTL启动子用于启动细胞膜上STL蛋白,通过调控应答途径进而控制甘油/H+进入细胞,使得产甘油假丝酵母更高效的表达。构建了含有gfp报告基因的重组表达载体pET-PSTL1/PSTL2-gfp-zeocin-rDNA,转入大肠杆菌中复制后进行验证;电击转化将线性片段pET-PSTL1/PSTL2-gfp-zeocin-rDNA转入产甘油假丝酵母,以抗性基因zeocin为筛选标记,培养后进行荧光检测发现转入后的荧光强度明显加强。实验结果表明,酿酒酵母的PSTL启动子使得产甘油假丝酵母更高效的表达,从而产生更高的经济效益。
This subject used the PSTL promoter of Saccharomyces cerevisiae to start the STL protein on the cell membrane,and then control the entry of glycerol/H+into the cell by regulating the response pathway,so as to make the expression of glycerol producing Candida more efficient.The recombinant expression vector pET-PSTL1/PSTL2-gfp-zeocin-rDNA containing GFP reporter gene is constructed and transferred into E.coli for replication.The linear fragment pET-PSTL1/PSTL2-gfp-zeocin rDNA is transferred into Candida glycerogens by electric shock transformation.The resistance gene zeocin is used as the screening marker.After cultured,the fluorescence intensity is significantly enhanced.The experimental results show that Saccharomyces cerevisiae PSTL promoter will enable more efficient expression of Candida glycerogenesis.so we can get higher economic benefits.
作者
杨海雁
于喆源
YANG Haiyan;YU Zheyuan(Zhangye Quality Inspection and Testing Institute,Gansu Zhangye 734000)
出处
《生物化工》
2021年第5期82-84,99,共4页
Biological Chemical Engineering
