Difference between revisions of "Part:BBa K3376004"

 
 
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<partinfo>BBa_K3376004 short</partinfo>
 
<partinfo>BBa_K3376004 short</partinfo>
  
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Thiol peroxidase of S. mutans plays a role in cell protection against oxidative stress by catalyzing the reduction of H2O2 to water. The gene expression is correlated to the concentration of H2O2. Thiol peroxidase promoter (tpxp) activity is induced by the presence of H2O2. The promoter was cloned out from the gDNA of S. mutans and tested in transformed E. coli.
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=== Expression ===
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The fluorescence expression levels were measured for lactate dehydrogenase promoter (ldhp) [ldhp-GFP-Tr/pSB1C3 (BBa_K3376002)] and thiol peroxidase promoter (tpxp) [tpxp-GFP-Tr/pSB1C3 (BBa_K3376004)] activity of S. mutans. GFP was detected at Ex/Em = 483/513. As shown in the figure, the expression level of tpxp-driven GFP was limited compared to a strong and constitutive promoter (ldhp).
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[[File:T--Mingdao--ww1.png|450px|center]]
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=== Regulation ===
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The tpxp-GFP-Tr/pSB1C3 [BBa_K3376002] and ldhp-AQP-Tr-tpxp-GFP-Tr/pSB1C3 [BBa_K3376007] were transformed into E. coli Nissle strain. Because of the limited H2O2 diffusion across the cell membrane, the tpxp activity was not changed in the presence of H2O2. In aid of expression of aquaporin (AQP) to facilitate H2O2 permeation, the tpxp-driven GFP levels were enhanced by the increasing amounts of H2O2 in a dose-dependent manner.
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[[File:T--Mingdao--ww3.png|600px|center]]
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<span class='h3bb'>Sequence and Features</span>
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=== <span class='h3bb'>Sequence and Features</span> ===
 
<partinfo>BBa_K3376004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3376004 SequenceAndFeatures</partinfo>
  

Latest revision as of 16:14, 6 October 2020


tpxp-GFP-Tr/pSB1C3

Thiol peroxidase of S. mutans plays a role in cell protection against oxidative stress by catalyzing the reduction of H2O2 to water. The gene expression is correlated to the concentration of H2O2. Thiol peroxidase promoter (tpxp) activity is induced by the presence of H2O2. The promoter was cloned out from the gDNA of S. mutans and tested in transformed E. coli.

Expression

The fluorescence expression levels were measured for lactate dehydrogenase promoter (ldhp) [ldhp-GFP-Tr/pSB1C3 (BBa_K3376002)] and thiol peroxidase promoter (tpxp) [tpxp-GFP-Tr/pSB1C3 (BBa_K3376004)] activity of S. mutans. GFP was detected at Ex/Em = 483/513. As shown in the figure, the expression level of tpxp-driven GFP was limited compared to a strong and constitutive promoter (ldhp).

T--Mingdao--ww1.png

Regulation

The tpxp-GFP-Tr/pSB1C3 [BBa_K3376002] and ldhp-AQP-Tr-tpxp-GFP-Tr/pSB1C3 [BBa_K3376007] were transformed into E. coli Nissle strain. Because of the limited H2O2 diffusion across the cell membrane, the tpxp activity was not changed in the presence of H2O2. In aid of expression of aquaporin (AQP) to facilitate H2O2 permeation, the tpxp-driven GFP levels were enhanced by the increasing amounts of H2O2 in a dose-dependent manner.

T--Mingdao--ww3.png


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 840