Difference between revisions of "Part:BBa K2936001"

 
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<partinfo>BBa_K2936001 short</partinfo>
 
<partinfo>BBa_K2936001 short</partinfo>
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The promoter is a part of the arabinose operon whose name derives from the genes it regulates transcription of: araB, araA, and araD. In E. coli, the PBAD promoter is adjacent to the PC promoter, which transcribes the araC gene in the opposite direction.
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The LacZ alpha gene is one of the structural genes present on the Lac Operon in E. coli bacteria . When transcribed by RNA polymerase , the LacZ alpha gene encoding β-galactosidase , an enzyme responsible for the hydrolysis of β-galactosides (e.g. 2-Nitrophenyl β-D-galactopyranoside (ONPG) will be expressed.
  
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When the AraBAD promoter activate, the LacZ&#593; expresses the &#946;-Gal and the culture with ONPG would turns orange.
 
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===Usage and Biology===
 
  
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===<h1>Usage and Biology</h1>===
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Figure 1 The A350 of reaction solution of bacterial culture and TMB
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Figure 2 The mechanism of the color reaction
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From figure 2, the color of the third group is enough obvious compared to the background. And at the same time, in order to balance the principle of economy, the ratio of bacterial culture (OD6000=0.5) to TMB is designed to be 1:10 .
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===Reference===
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[1]Jesus Fernandez-Rodriguez, Felix Moser, Miryoung Song, Christopher A Voigt, (2017) Engineering RGB color vision into Escherichia coli. [J]. Biol. Chem. 13,706-710
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 06:25, 19 October 2019


pBAD-LacZɑ

The promoter is a part of the arabinose operon whose name derives from the genes it regulates transcription of: araB, araA, and araD. In E. coli, the PBAD promoter is adjacent to the PC promoter, which transcribes the araC gene in the opposite direction.

The LacZ alpha gene is one of the structural genes present on the Lac Operon in E. coli bacteria . When transcribed by RNA polymerase , the LacZ alpha gene encoding β-galactosidase , an enzyme responsible for the hydrolysis of β-galactosides (e.g. 2-Nitrophenyl β-D-galactopyranoside (ONPG) will be expressed.


Usage and Biology


Figure 1 The A350 of reaction solution of bacterial culture and TMB
Figure 2 The mechanism of the color reaction
From figure 2, the color of the third group is enough obvious compared to the background. And at the same time, in order to balance the principle of economy, the ratio of bacterial culture (OD6000=0.5) to TMB is designed to be 1:10 .

Reference

[1]Jesus Fernandez-Rodriguez, Felix Moser, Miryoung Song, Christopher A Voigt, (2017) Engineering RGB color vision into Escherichia coli. [J]. Biol. Chem. 13,706-710 Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 120
    Illegal BamHI site found at 182
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]