Difference between revisions of "Part:BBa K3790085"
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Gp2 is an early phage gene, which means they are transcribed by the host's RNA polymerase at the very beginning of the infection process<ref>Roles of the early genes of bacteriophage T7 in shutoff of host macromolecular synthesis. McAllister WT,  Barrett CL. J Virol, 1977 Sep;23(3):543-53.  PMID:330878</ref>.  
 
Gp2 is an early phage gene, which means they are transcribed by the host's RNA polymerase at the very beginning of the infection process<ref>Roles of the early genes of bacteriophage T7 in shutoff of host macromolecular synthesis. McAllister WT,  Barrett CL. J Virol, 1977 Sep;23(3):543-53.  PMID:330878</ref>.  
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Gp2 is a small protein that can bind to both the 1.1 domain of σ70 factor and the β’ subunit of the host's RNAP<ref>Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2. James E,  Liu M,  Sheppard C,  Mekler V,  Cámara B,  Liu B,  Simpson P,  Cota E,  Severinov K,  Matthews S,  Wigneshweraraj S. Mol Cell, 2012 Sep 14;47(5):755-66.  PMID:22819324
 
Gp2 is a small protein that can bind to both the 1.1 domain of σ70 factor and the β’ subunit of the host's RNAP<ref>Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2. James E,  Liu M,  Sheppard C,  Mekler V,  Cámara B,  Liu B,  Simpson P,  Cota E,  Severinov K,  Matthews S,  Wigneshweraraj S. Mol Cell, 2012 Sep 14;47(5):755-66.  PMID:22819324
 
</ref>, results in effective inhibiting of the transcription initiated by σ70-RNAP complex.
 
</ref>, results in effective inhibiting of the transcription initiated by σ70-RNAP complex.
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This part can be used to produce gp2 after arabinose induction.
  
 
===Experimental Results===
 
===Experimental Results===
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Gp2 was eliminated by us because gp5.7 is a better choice. For more details, please visit: https://parts.igem.org/wiki/index.php?title=Part:BBa_K3790050
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Gp2 was eliminated by us because gp5.7 is a better choice. For detailed reasons, please visit:
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* https://parts.igem.org/Part:BBa_K3790232
  
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[[File:T--Fudan--K3790231fig1.png|none|600px|thumb|Figure 1. Growth curve of strains with pBAD driven gp2 under different induction level.]]
  
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[[File:T--Fudan--K3790231fig2.png|none|600px|thumb|Figure 2. GFP/OD600 curve of strains with pBAD driven gp2 under different induction level. The GFP is σ70-driven.]]
  
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===Reference===
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From these two figures, we can conclude that arabinose induction level can't notably influence the effects of pBAD driven gp2 on inhibiting transcription initiated by σ70-RNAP complex and on hindering cell growth.
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<references />
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'>'''Sequence and Features'''</span>
 
<partinfo>BBa_K3790085 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3790085 SequenceAndFeatures</partinfo>
  
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<partinfo>BBa_K3790085 parameters</partinfo>
 
<partinfo>BBa_K3790085 parameters</partinfo>
 
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===Reference===
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<references />

Latest revision as of 04:00, 22 October 2021


T7 phage gene product 2, an E.coli transcription inhibitor


Introduction

2021 Fudan

In order to turn the host bacteria into ‘factories’ that concentrate on manufacturing parts of the virus, some phages have the ability to shut off the endogenous transcription of host bacteria. As for T7 phage, there are three gene products collaboratively realizing the function, which are gene product (gp) 0.7, gp2 and gp5.7[1].

Gp2 is an early phage gene, which means they are transcribed by the host's RNA polymerase at the very beginning of the infection process[2].



Usage and Biology

Gp2 is a small protein that can bind to both the 1.1 domain of σ70 factor and the β’ subunit of the host's RNAP[3], results in effective inhibiting of the transcription initiated by σ70-RNAP complex.

This part can be used to produce gp2 after arabinose induction.

Experimental Results

Gp2 was eliminated by us because gp5.7 is a better choice. For detailed reasons, please visit:

Figure 1. Growth curve of strains with pBAD driven gp2 under different induction level.
Figure 2. GFP/OD600 curve of strains with pBAD driven gp2 under different induction level. The GFP is σ70-driven.

From these two figures, we can conclude that arabinose induction level can't notably influence the effects of pBAD driven gp2 on inhibiting transcription initiated by σ70-RNAP complex and on hindering cell growth.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Reference

  1. ↑ T7 phage factor required for managing RpoS in Escherichia coli. Tabib-Salazar A,  Liu B,  Barker D,  Burchell L,  Qimron U,  Matthews SJ,  Wigneshweraraj S. Proc Natl Acad Sci U S A, 2018 Jun 5;115(23):E5353-E5362. PMID:29789383
  2. ↑ Roles of the early genes of bacteriophage T7 in shutoff of host macromolecular synthesis. McAllister WT,  Barrett CL. J Virol, 1977 Sep;23(3):543-53. PMID:330878
  3. ↑ Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2. James E,  Liu M,  Sheppard C,  Mekler V,  Cámara B,  Liu B,  Simpson P,  Cota E,  Severinov K,  Matthews S,  Wigneshweraraj S. Mol Cell, 2012 Sep 14;47(5):755-66. PMID:22819324