Difference between revisions of "Part:BBa K2997003"
 
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<partinfo>BBa_K2997003 short</partinfo>
 
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The construction and digestion of DNA gel shows that the size of the pchR-GFP composite part was as expected.
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===Background===
  
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The pchR-GFP composite part (BBa_K2997008) was constructed from IDT DNA synthesis. After undergoing a few rounds of overlap PCR to combine 3 separate fragments into one long composite part, the part is then inserted into pSB1C3 and electroporated into E. coli Top 10. After allowing the transformed E. coli Top 10 to grow on LB+Cm plates, colony PCR and sequencing was carried out to determine the clone we got was positive or not. The colony PCR results indicated that the E. coli has pSB1C3 with pchR-GFP, but, the sequencing results indicated that random mutations have occurred on the pchR-GFP composite part.  
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The <i>pchR</i> gene is responsible for the production of activator proteins that can bind with the inducer (<i>p</i>-Cresol) and leads to the conformation of the protein. <sup>[1][2]</sup> In other words, the activator protein can bind with <i>p</i>-Cresol to form a protein-<i>p</i>-Cresol complex that will then bind onto the <i>p</i>-Cresol sensing region located just downstream of the <i>pchR</i> gene. The binding of the protein-<i>p</i>-Cresol complex initiates the expression of the downstream gene, in our case GFP.
  
  
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<partinfo>BBa_K2997003 parameters</partinfo>
 
<partinfo>BBa_K2997003 parameters</partinfo>
 
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===References===
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<li>Cho, Ah Ra, Eun Jin Lim, Yaligara Veeranagouda, and Kyoung Lee* Department of Microbiology, Changwon National University, Changwon-si, Kyongnam 641-773, Korea . Identification of a <i>p</i>-Cresol Degradation Pathway by a GFP-Based Transposon in <i>Pseudomonas</i> and Its Dominant Expression in Colonies</li>
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<li>Merike Jo ˜esaar, Eeva Heinaru, Signe Viggor, Eve Vedler & Ain Heinaru Institute of Molecular and Cell Biology, Tartu University, Tartu, Estonia. Diversity of the transcriptional regulation of the pch gene cluster in two indigenous <i>p</i>-Cresol-degradative strains of <i>Pseudomonas fluorescens</i>
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Latest revision as of 07:15, 20 October 2019


Sigma 54-dependent transcriptional regulator (pchR)

Background

The pchR gene is responsible for the production of activator proteins that can bind with the inducer (p-Cresol) and leads to the conformation of the protein. [1][2] In other words, the activator protein can bind with p-Cresol to form a protein-p-Cresol complex that will then bind onto the p-Cresol sensing region located just downstream of the pchR gene. The binding of the protein-p-Cresol complex initiates the expression of the downstream gene, in our case GFP.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1738
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1151
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1196
    Illegal AgeI site found at 179
    Illegal AgeI site found at 1079
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1105


References


  1. Cho, Ah Ra, Eun Jin Lim, Yaligara Veeranagouda, and Kyoung Lee* Department of Microbiology, Changwon National University, Changwon-si, Kyongnam 641-773, Korea . Identification of a p-Cresol Degradation Pathway by a GFP-Based Transposon in Pseudomonas and Its Dominant Expression in Colonies
  2. Merike Jo ˜esaar, Eeva Heinaru, Signe Viggor, Eve Vedler & Ain Heinaru Institute of Molecular and Cell Biology, Tartu University, Tartu, Estonia. Diversity of the transcriptional regulation of the pch gene cluster in two indigenous p-Cresol-degradative strains of Pseudomonas fluorescens