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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</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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It can be used to inhibit E.coli. endogenous transcription and hinder E.coli. growth.
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This part can be used to produce gp2 after arabinose induction.
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iGEM16_Imperial_College has registered a so-called T7 gp2 (BBa_K1893019), but strangely, it's not the same thing as the T7 gp2 we use, whose coding sequence is got from NCBI, and it's true that the attributes of their gp2 are totally different from ours.
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===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/Part:BBa_K3790050
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Gp2 was eliminated by us because gp5.7 is a better choice. For more details, please visit:
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* https://parts.igem.org/Part:BBa_K3790050
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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--K3790231fig1.png|none|600px|thumb|Figure 1. No Time to Write a Title. We will present the data in our video.]]
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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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[[File:T--Fudan--K3790231fig2.png|none|600px|thumb|Figure 2. No Time to Write a Title. We will present the data in our video.]]
 
  
 
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Revision as of 03:18, 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 more details, 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.


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