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Related Concept Videos

Repressible Operon: trp Operon01:21

Repressible Operon: trp Operon

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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...
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Constitutive and Regulated Gene Expression01:27

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Gene expression in prokaryotes is governed by constitutive and regulated systems, allowing cells to balance the production of essential proteins with adaptive responses to environmental changes.Constitutive Gene ExpressionConstitutive, or housekeeping, genes are continuously expressed as they encode proteins vital for fundamental cellular processes. These include enzymes for glycolysis, ribosomal components for protein synthesis, and proteins involved in DNA replication. Their constant...
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Prokaryotic Transcriptional Activators and Repressors01:58

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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.
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The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
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Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
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What is Gene Expression?01:42

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Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
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Sealable Femtoliter Chamber Arrays for Cell-free Biology
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A Temperature-Controlled Cell-Free Expression System by Dynamic Repressor.

Junzhu Yang1, Chen Wang1, Yuan Lu1

  • 1Key Laboratory of Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.

ACS Synthetic Biology
|March 23, 2022
PubMed
Summary
This summary is machine-generated.

This study introduces a temperature-controlled cell-free protein synthesis (tcCFPS) system for precise control over protein production. The novel tcCFPS system demonstrates a significant 143-fold dynamic range in protein expression, enhancing synthetic biology applications.

Keywords:
artificial cellcI repressorcell-free protein synthesiscell-free systemthermal switch

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Area of Science:

  • Synthetic biology
  • Biochemistry
  • Molecular biology

Background:

  • Cell-free protein synthesis (CFPS) is a key platform in synthetic biology for protein production.
  • Traditional CFPS methods face limitations in achieving precise spatiotemporal control over protein synthesis.
  • Chemical induction methods can cause undesirable side effects.

Purpose of the Study:

  • To design and implement a temperature-controlled cell-free protein synthesis (tcCFPS) system for enhanced control.
  • To utilize the repressor cI protein from E. coli for temperature-based regulation.
  • To explore the application of tcCFPS in artificial cell systems.

Main Methods:

  • Development of a tcCFPS system based on E. coli.
  • Incorporation of the repressor cI protein for temperature-dependent gene expression control.
  • Construction of temperature-controlled artificial cells utilizing the tcCFPS system.

Main Results:

  • The tcCFPS system achieved a 143-fold dynamic range in protein expression levels between 30 °C and 37 °C.
  • Demonstrated precise spatiotemporal control over protein synthesis using temperature as an inducer.
  • Successfully constructed artificial cells regulated by temperature, expanding tcCFPS applications.

Conclusions:

  • Temperature serves as an effective and minimally invasive control switch for cell-free protein synthesis.
  • The developed tcCFPS system offers a robust method for active protein synthesis in cell-free environments.
  • This technology holds potential for applications in drug synthesis and targeted delivery systems.