Difference between revisions of "Part:BBa K2996703"

 
 
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Cas1 and Cas2 are the only Cas proteins required for genetic recording of infections through spacer acquisition from invader DNA, thus they are called adaptation complex. In adaptation stage, short DNA fragments generated from phage DNA or other foreign DNA are captured by the adaptation complex. Following trimming of excess DNA, the adaptation complex integrates these fragments into the CRISPR array.  
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RpoA encodes the DNA-directed RNA polymerase subunit alpha, which plays an important role in subunit assembly since its dimerization is the first step in the sequential assembly of subunits to form the holoenzyme. It may activate transcription more effectively, compared to RpoZ used in other experiments.  
  
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This RpoA-dCas9 fusion protein should be an innovated transcription activator.
  
  
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===Background===
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===Usage and Biology===
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<center>{{#tag:html|<img style="max-width: 50%" src="https://2019.igem.org/wiki/images/d/d0/T--SJTU-BioX-Shanghai--RFP4.png" alt="" />}}</center>
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<center><b>Figure 1.</b> <i>Schematic diagram of RpoA and transcription activation</i> </center>
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DNA-directed RNA polymerases are responsible for the polymerization of ribonucleotides into a sequence complementary to the template DNA.
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Most RNA polymerases are multimeric enzymes and are composed of a variable number of subunits. The core RNA polymerase complex consists of five subunits (two alpha, one beta, one beta-prime and one omega) and is sufficient for transcription elongation and termination but is unable to initiate transcription. Transcription initiation from promoter elements requires a sixth, dissociable subunit called a sigma factor, which reversibly associates with the core RNA polymerase complex to form a holoenzyme. The core RNA polymerase complex forms a "crab claw"-like structure with an internal channel running along the full length. The key functional sites of the enzyme, as defined by mutational and cross-linking analysis, are located on the inner wall of this channel.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2996703 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K2996703 parameters</partinfo>
 
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Latest revision as of 23:29, 21 October 2019


RpoA, DNA-directed RNA polymerase subunit alpha

RpoA encodes the DNA-directed RNA polymerase subunit alpha, which plays an important role in subunit assembly since its dimerization is the first step in the sequential assembly of subunits to form the holoenzyme. It may activate transcription more effectively, compared to RpoZ used in other experiments.

This RpoA-dCas9 fusion protein should be an innovated transcription activator.


Background

Figure 1. Schematic diagram of RpoA and transcription activation

DNA-directed RNA polymerases are responsible for the polymerization of ribonucleotides into a sequence complementary to the template DNA. Most RNA polymerases are multimeric enzymes and are composed of a variable number of subunits. The core RNA polymerase complex consists of five subunits (two alpha, one beta, one beta-prime and one omega) and is sufficient for transcription elongation and termination but is unable to initiate transcription. Transcription initiation from promoter elements requires a sixth, dissociable subunit called a sigma factor, which reversibly associates with the core RNA polymerase complex to form a holoenzyme. The core RNA polymerase complex forms a "crab claw"-like structure with an internal channel running along the full length. The key functional sites of the enzyme, as defined by mutational and cross-linking analysis, are located on the inner wall of this channel.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 459
  • 1000
    COMPATIBLE WITH RFC[1000]