摘 要：在转录的过程中，控制转录因子（TFs）可以对转录进行调控，由于TFs能够指定RNA聚合酶全酶和启动子的结合，因此可以以此来决定转录的过程。通过定向进化（或合理设计）中获得的突变体来替换内源性的TFs，以及引入兼容的异构TFs，是有效的调节转录的途径。同时遵循蛋白质的设计原理，设计具有预期功能的人工TFs，并利用从进化的角度对相关转录因子的多重序列比对中得到的信息来重建祖先转录因子，也是一种可行的代替方案。诱导蛋白表达系统可以由工程改造后的转录因子来创建，以此来对特定的刺激做出响应。此外，改造后的转录因子可以引导一种不同于野生型菌株的转录模式。因此，改造后的转录因子可以被用于构建耐不良条件的菌株，而由于耐药性的出现通常需要全部转录在转录组测序中做出改变。与此同时，这些改造后的转录因子也能被用于构建复杂设计的基因回路以达到不同的用途。本篇就上述内容的研究进展进行综述。

关键词：转录因子(TF)，蛋白质工程，转录组，全局调控，祖先序列重建，基因回路，高通量筛选(HTS)

Abstract：Transcription can be regulated by controlling the provision of transcriptional factors (TFs),because TFs determine the transcription process by specifying the binding of RNA polymerase holoenzyme on promoters. Manipulations, such as replacing endogenous TFs with mutants obtainedthrough directed evolution (or rational design),and introducing compatible heterogeneous TFs, are effective ways to regulate transcription. Designing artificial TFs with desired properties by following the principles of protein design, and reconstructing ancestral TFs with the information gained from a perspective of evolution by multiple sequence alignments of the related TFs, are feasible alternative options. The engineered TFs can be used to create inducible protein expression systems that respond to a specific stimulus. Moreover, the engineered TFs could lead to a transcriptional profile different from that of the wild-type strains. Accordingly, these engineered TFs could be utilized to construct strainsresistant to adverse conditions,since the emergence of resistance generally requires global transcriptional changes at the transcriptomic scale. Meanwhile, these engineered TFs can also be employed in the construction of sophisticatedly design genetic circuits for diverse purposes. This paper reviews the research progress of the above content.

Keywords：Transcriptional factor (TF)；Protein engineering；Transcriptome；Global regulation；Ancestral sequence reconstruction；Genetic circuit；high-throughput screening