在rhaBAD促进体的控制下,代码子优化的Taq DNA聚合酶的高水平表达
Fina Amreta Laksmi1, Kartika Sari Dewi2, Isa Nuryana1
1Research Center for Applied Microbiology, National Research and Innovation Agency, Jalan Raya Bogor KM 46, Cibinong, Bogor, 16911, West Java, Indonesia.
Analytical biochemistry
|May 30, 2024
概括
研究人员利用rhaBAD促进剂优化了大肠杆菌中Taq DNA聚合酶的产生. 这种N终端删除策略实现了表达的3倍增加,产生8.5mg/L的纯化酶.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 酶工程是指酶工程的工程.
背景情况:
- 来自Thermus aquaticus的Taq DNA聚合酶对于聚合酶链反应 (PCR) 应用至关重要.
- 高商业需求需要Taq DNA聚合酶的高效生产方法.
研究的目的:
- 开发E. coli中Taq DNA聚合酶的高产量的表达系统.
- 使用可控制的促进剂优化Taq DNA聚合酶N端删除突变的产生.
主要方法:
- 利用l-rhamnose诱导的rhaBAD促进物用于控制大肠杆菌中的重组蛋白表达.
- 设计了Taq DNA聚合酶的N端删除.
- 优化了子使用和培养条件,以提高蛋白质表达水平.
主要成果:
- 达到了Taq DNA聚合酶表达的显著改善,高达基线的3倍.
- 成功地产生了大约8.5mg/L的净化N端删除的Taq DNA聚合酶.
- 证明了rhaBAD促进体系统在高水平重组蛋白质生产中的有效性.
结论:
- 在大肠杆菌中,Taq DNA聚合酶的N终端删除突变可以使用rhaBAD促进体有效地产生.
- 子优化和培养条件精制是提高重组酶产量的有效策略.
- 这项研究提出了一种新的方法,用于生产高纯度Taq DNA聚合酶,用于商业应用.
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