Toxic protein expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system
Toxic protein expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system
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DOI：
10.2144/000112112
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发表时间：
2006-03-01
期刊：
影响因子：
2.7
通讯作者：
Surber, MW
中科院分区：
文献类型：
--
作者：
Giacalone, MJ;Gentile, AM;Surber, MW
严格调控的原核表达系统的改进允许有毒重组蛋白的功能表达是一个不断进化的过程。遗憾的是，目前最好的启动子选择要么被紧密抑制并产生少量蛋白质，要么产生大量蛋白质但缺乏必要的抑制以避免在没有诱导剂的情况下因泄漏表达而引起的突变。在本报告中，我们提出了三种新的受L严格调控的原核表达结构-鼠李糖和D-葡萄糖。这些表达载体利用大肠杆菌RHAT启动子和相应的调控基因来提供可滴定的、高水平的蛋白质产量，而不影响克隆的完整性。这些成分结合在一起，可能会使其他有毒蛋白的稳定克隆和功能表达成为可能。
The refinement of tightly regulated prokaryotic expression systems that permit functional expression of toxic recombinant proteins is a continually evolving process. Unfortunately, the current best promoter options are either tightly repressed and produce little protein, or produce substantial protein but lack the necessary repression to avoid mutations stimulated by leaky expression in the absence of inducer In this report, we present three novel prokaryotic expression constructs that are tightly regulated by L-rhamnose and D-glucose. These expression vectors utilize the Escherichia coli rhaT promoter and corresponding regulatory genes to provide titratable, high-level protein yield without compromising clone integrity. Together these components may enable the stable cloning and functional expression of otherwise toxic proteins.