Difference between revisions of "Part:BBa K2955002"

 
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<partinfo>BBa_K2955002 short</partinfo>
 
<partinfo>BBa_K2955002 short</partinfo>
  
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glpB is a gene that was found to be related to degradation of glyphosate in the bacterium Burholderia psuedomallei (formerly Psuedomonas). This gene was utilized as part of a two gene system along with glpA (BBa_K2955001) with the overall goal of degrading glyphosate. This gene was reported by Penaloza-Vazquez et al (1994) as allowing glyphosate to be used as a phosphorous source by breaking the C-N bond.  
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glpB is a gene that was found to be related to degradation of glyphosate in the bacterium Burkholderia psuedomallei (formerly Psuedomonas). This gene was utilized as part of a two gene system along with glpA (BBa_K2955001) with the overall goal of degrading glyphosate. This gene was reported by Penaloza-Vazquez et al (1994) as allowing glyphosate to be used as a phosphorous source by breaking the C-N bond.  
  
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===Usage and Biology===
 
===Usage and Biology===
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glpB was used as part of a two part system with the goal of increasing cell survivability and degradation of glyphosate. glpB is associated with the breakdown of glyphosate through the glyphosate oxidoreductase pathway, in which the glyphosate molecule is broken at the C-N bond to form aminomethylphosphonic acid (AMPA) and glyoxylate. In our system, glpB was regulated by pLac, and in experiments expression was induced by adding IPTG.
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==Methods==
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This part was tested by running growth curves under various conditions. Growth curve data was collected using a Bioscreener, Bioscreen C. Overnight cultures of the full plasmid, which had both glpA and glpB (BBa_K2955002), and a control culture were grown overnight in tubes of LB+chloramphenicol at 37C and 220 RPM. 3.0 mL of the tested concentrations of pure glyphosate and Roundup were prepared with 3 uL of chloramphenicol. IPTG was added to reach a final concentration of 0.5 mM IPTG in induced samples.Each concentration was tested with 4 replicates. Each well was inoculated with 3 uL of cells and 300 uL of the stock concentrations. The plates were left in the Bioscreener for 96 hours at 37 C and meadium shaking. Measurements were taken every 30 minutes. It was not possible to test glpB independently of glpA due to the constitutive promoter regulating glpA. The following data is taken in comparison to Rounddown when glpB is not expressed.
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==Results==
  
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K2955002 parameters</partinfo>
 
<partinfo>BBa_K2955002 parameters</partinfo>
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<!-- -->Burkholderia

Revision as of 07:56, 21 October 2019


glpB

glpB is a gene that was found to be related to degradation of glyphosate in the bacterium Burkholderia psuedomallei (formerly Psuedomonas). This gene was utilized as part of a two gene system along with glpA (BBa_K2955001) with the overall goal of degrading glyphosate. This gene was reported by Penaloza-Vazquez et al (1994) as allowing glyphosate to be used as a phosphorous source by breaking the C-N bond.

Usage and Biology

glpB was used as part of a two part system with the goal of increasing cell survivability and degradation of glyphosate. glpB is associated with the breakdown of glyphosate through the glyphosate oxidoreductase pathway, in which the glyphosate molecule is broken at the C-N bond to form aminomethylphosphonic acid (AMPA) and glyoxylate. In our system, glpB was regulated by pLac, and in experiments expression was induced by adding IPTG.

Methods

This part was tested by running growth curves under various conditions. Growth curve data was collected using a Bioscreener, Bioscreen C. Overnight cultures of the full plasmid, which had both glpA and glpB (BBa_K2955002), and a control culture were grown overnight in tubes of LB+chloramphenicol at 37C and 220 RPM. 3.0 mL of the tested concentrations of pure glyphosate and Roundup were prepared with 3 uL of chloramphenicol. IPTG was added to reach a final concentration of 0.5 mM IPTG in induced samples.Each concentration was tested with 4 replicates. Each well was inoculated with 3 uL of cells and 300 uL of the stock concentrations. The plates were left in the Bioscreener for 96 hours at 37 C and meadium shaking. Measurements were taken every 30 minutes. It was not possible to test glpB independently of glpA due to the constitutive promoter regulating glpA. The following data is taken in comparison to Rounddown when glpB is not expressed.

Results

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 147
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Burkholderia