生物技术通报 ›› 2017, Vol. 33 ›› Issue (1): 120-128.doi: 10.13560/j.cnki.biotech.bull.1985.2017.01.013

• 研究报告 • 上一篇    下一篇

酿酒酵母人工杂合启动子与天然启动子活性比较

唐瑞琪1, 熊亮2, 白凤武1, 2, 赵心清1   

  1. 1. 上海交通大学生命科学技术学院,上海 200240;
    2. 大连理工大学生命科学与技术学院,大连 116023
  • 收稿日期:2016-10-20 出版日期:2017-01-25 发布日期:2017-01-19
  • 作者简介:唐瑞琪,女,博士研究生,研究方向:酿酒酵母代谢工程改造;E-mail:rq_tang@sjtu.edu.cn
  • 基金资助:
    国家自然科学基金项目(31461143029,5151101168)

Activity Comparison of the Artificial Hybrid Promoter with Its Native Promoter in Saccharomyces cerevisiae

TANG Rui-qi1, XIONG Liang2, BAI Feng-wu1, 2, ZHAO Xin-qing1   

  1. 1. School of Life Science and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240;
    2. School of Life Science and Biotechnology,Dalian University of Technology,Dalian 116023
  • Received:2016-10-20 Published:2017-01-25 Online:2017-01-19

摘要: 系统地比较了人工启动子PaTEF1与天然启动子PTEF1和PTDH3在不同条件下的活性差别,结果表明,人工启动子PaTEF1的活性并不是在任何条件下都高于天然启动子的活性,而与宿主、培养基以及细胞生长阶段有关。在3个宿主背景中,BY4741中PaTEF1的活性最高,而LX03中最低。在YPD100中,人工启动子PaTEF1活性分别为天然启动子PTEF1和PTDH3活性的1.4-4.6倍和0.9-2.0倍;而在YPE(5%和7%)中,PaTEF1活性与PTEF1和PTDH3活性之比在0.7-1.3以及0.8-1.3之间。在YPE中培养时,PaTEF1的活性为在YPD100中培养时的1.7-2.0倍,PTEF1和PTDH3的活性在YPE中为YPD100中培养时的2.7-7.1倍和1.3-3.4倍,启动子在YPE中的活性较YPD100中更高,但人工启动子的活性变化较天然启动子更小。此外,在不同遗传背景的菌株中,启动子活性从对数期早期到对数期中期和从对数期中期到稳定期的变化趋势不同。

关键词: 酿酒酵母, 人工杂合启动子, TEF1启动子, 宿主菌株, 环境条件

Abstract: The responses of promoter strengths of artificial TEF1 promoter(PaTEF1)and native promoter PTEF1 as well as PTDH3 were comprehensively compared. The strength of PaTEF1 was not always higher than PTEF1,but varied along with the genetic background of host,medium,and cell growth phase. Among the three investigated hosts,PaTEF1 showed the highest strength in BY4741,while the lowest in LX03. The strengths of PaTEF1 were 1.4-4.6 and 0.9-2.0 times of those of native promoters(PTEF1 and PTDH3)in YPD100 medium,respectively. The activities of PaTEF1 in YPE(5% and 7%)were 0.7-1.3 and 0.8-1.3 times of PTEF1 and PTDH3. The activity of PaTEF1 in YPE was 1.7-2.0 times of that in YPD100,whereas 2.7-7.1 and 1.3-3.4 times for PTEF1 and PTDH3,respectively. The activities of the promoters in YEP were higher than those in YPD100,however,the activity variation of artificial promoters was less than that of native promoter. In addition,the variation trends of promoter strengths from early-log to mid-log phase and from mid-log to stationary phase varied in different host strains.

Key words: Saccharomyces cerevisiae, artificial hybrid promoter, TEF1 promoter, host strain, environmental condition