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Related Concept Videos

Transcription01:10

Transcription

157.0K
Overview
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
157.0K
Transcription Attenuation in Prokaryotes02:42

Transcription Attenuation in Prokaryotes

18.5K
Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
18.5K
Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

27.1K
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
27.1K
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

32.8K
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
32.8K
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

11.2K
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
11.2K
Prokaryotic Transcriptional Activators and Repressors01:58

Prokaryotic Transcriptional Activators and Repressors

25.5K
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...
25.5K

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XNA Synthesis and Reverse Transcription by Engineered Thermophilic Polymerases.

Christopher Cozens1, Vitor B Pinheiro1,2

  • 1Institute for Structural and Molecular Biology, Division of Biosciences, University College London, London, United Kingdom.

Current Protocols in Chemical Biology
|July 25, 2018
PubMed
Summary

Researchers engineered a Thermococcus gorgonarius polymerase to synthesize hexitol nucleic acid (HNA) and other xenonucleic acids (XNAs). This protocol enables large-scale XNA synthesis and characterization for diverse biotechnological applications.

Keywords:
XNA reverse transcriptionXNA synthesisthermostable DNA polymerases

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Synthetic Biology

Background:

  • Hyperthermophilic archaeal B-family polymerases are versatile for engineering extended substrate ranges.
  • These enzymes can synthesize and reverse transcribe various xenonucleic acids (XNAs) distinct from canonical DNA.

Purpose of the Study:

  • To present a protocol for hexitol nucleic acid (HNA) synthesis using an engineered Thermococcus gorgonarius polymerase variant.
  • To adapt the protocol for large-scale synthesis, purification, and characterization of XNAs.

Main Methods:

  • Engineering of Thermococcus gorgonarius polymerase for XNA synthesis.
  • Development of protocols for large-scale HNA synthesis and purification.
  • Adaptation of XNA purification and reverse transcription methods.

Main Results:

  • Demonstrated efficient synthesis of HNA by the engineered polymerase.
  • Established methods for large-scale HNA production and purification.
  • Provided insights into XNA reaction characterization and optimization.

Conclusions:

  • Engineered archaeal polymerases provide a robust platform for XNA synthesis.
  • The presented protocol facilitates scalable production and application of HNA and other XNAs.
  • This work advances the field of synthetic nucleic acids and their potential uses.