摘要
利用PCR技术从酿酒酵母基因组克隆得到甘油代谢关键酶基因gpd1启动子,并成功构建真核生物穿梭表达载体pYX212-zoecin-PScgpd1-GUS,并将其电击转入酿酒酵母中。将构建成功的酿酒酵母Saccharomyces cerevisiae基因工程菌分别在0.2,0.5,1.0 mol/L NaCl的盐胁迫下培养,首次通过GUS组织化学染色法和荧光法测定GUS报告基因的瞬时表达酶活检测gpd1启动子的酶活表达。研究发现,酵母甘油代谢关键酶基因gpd1启动子在不同渗透压下的表达有明显的差异。证实了gpd1启动子是受渗透压调节的,属于诱导型启动子,这可能与渗透压胁迫下的甘油代谢密切关联,相关研究未见报道。
A shuttle vector pYX212-zeocin-PScgpdl-GUS had been constructed and transformed into Saccharomyces cerevisiae by electroporation. The transformants was cultured in medium with 0.2, 0.5 and 1.0 mol/L NaCl supplement, respectively. The enzyme activity of promoter gpdl was determined by transient histochemical analysis of GUS gene at different osmotic pressure. The result showed that the expression of promoter gpdl in S. cerevisiae at different osmotic pressure were prominently different. It was concluded that the promoter gpdl was an inducible promoter, regulated by osmotic pressure.
出处
《食品与生物技术学报》
CAS
CSCD
北大核心
2008年第5期27-32,共6页
Journal of Food Science and Biotechnology
基金
国家自然科学基金项目(30570142
20676053)
国家863计划项目(2006AA020103)
江苏省青年科技创新人才基金项目(BK2006504)
长江学者和创新团队发展计划项目(IRT0532)
