在2种变体上对环素运营者的环素抑制器功能的高通量表征
Alexa N Gormick1, Adam M Zahm1, Samuel R Himes1
1Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112, United States.
ACS synthetic biology
|June 9, 2025
概括
研究人员设计了一种新的记者系统,以改善四环素诱导基因表达. 这个系统允许精细调整,动态控制哺乳动物细胞中的基因转录,推进细胞工程.
科学领域:
- 分子生物学分子生物学
- 细胞工程 细胞工程
- 系统生物学 系统生物学
背景情况:
- 环素诱导系统对于细胞工程中的转基因活动调节至关重要.
- 这些系统提供选择性,高亲和力和诱导性,但仅限于二进制开/关表达.
- 这种二元性质限制了复杂的监管系统和生物建模中的应用.
研究的目的:
- 开发一种高通量报告系统,以优化四环素诱导基因表达.
- 为了研究四环素耐药性经营者变体的和突变性库.
- 为了实现对哺乳动物细胞基因表达的动态和模块化控制.
主要方法:
- 开发一个高通量报告系统.
- 调查四环素耐药性经营者变体的和突变性库.
- 在哺乳动物细胞的单核酸分辨率下绘制Tet抑制剂DNA结合蛋白的功能相互作用.
主要成果:
- 确定了一系列变异效应,从抑制损失到自然操作者水平.
- 在单核酸分辨率下绘制了Tet抑制剂的功能相互作用.
- 坐标测定验证了观察到的变异效应.
结论:
- 开发的报告员系统使得对四环素耐药性的操作者序列特异性的全面表征成为可能.
- 这便于构建可变抑制,可诱导的系统.
- 能够对哺乳动物细胞培养中的基因表达进行动态和模块化控制.
相关概念视频
Repressible Operon: trp Operon
174
The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
174
Prokaryotic Transcriptional Activators and Repressors
22.3K
The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
Transcription of prokaryotic...
Transcription of prokaryotic...
22.3K
Operon Model
130
The operon model represents a fundamental mechanism of gene regulation in prokaryotes, enabling coordinated expression of genes involved in related metabolic or functional pathways. Operons consist of structural genes, a promoter, and an operator, with transcription regulated by repressors, activators, and small effector molecules.Structure and Function of OperonsAn operon is a cluster of structural genes transcribed together under the control of a single promoter. The promoter region...
130
Inducible Operons: lac Operon
179
The lac operon in Escherichia coli is a model for understanding inducible gene regulation and metabolic flexibility. It integrates local control by lactose and global regulation through catabolite repression, enabling E. coli to preferentially metabolize glucose when available and switch to lactose utilization when glucose is scarce.Structure and Function of the lac OperonThe lac operon contains three structural genes: lacZ (β-galactosidase), lacY (lactose permease), and lacA...
179
Operons
49.9K
Prokaryotes can control gene expression through operons—DNA sequences consisting of regulatory elements and clustered, functionally related protein-coding genes. Operons use a single promoter sequence to initiate transcription of a gene cluster (i.e., a group of structural genes) into a single mRNA molecule. The terminator sequence ends transcription. An operator sequence, located between the promoter and structural genes, prohibits the operon’s transcriptional activity if bound by...
49.9K


