Difference between revisions of "Part:BBa K2325102"
 
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<partinfo>BBa_K2325102 short</partinfo>
 
<partinfo>BBa_K2325102 short</partinfo>
  
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The BBa_K2325101 contains formamidase gene with T7 promoter and terminator.
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The BBa_K2325102 contains phosphite dehydrogenase gene with T7 promoter and terminator.
  
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===Usage and Biology===
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Phosphite dehydrogenase is an enzyme involved in the production of phosphate for providing phosphorus source for the engineering bacteria. This part has been constructed into expression vectors to modify the metabolic pathway of phosphorus in E.coli.
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<center>
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[[File:BBa_K2325102--1.png|750px]]<br />
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<b>Figure 1. Growth curve of <i>E.coli</i> BL21(DE3) and the engineering <i>E.coli</i> BL21(DE3)(pGEX-ptx) expressing phosphite dehydrogenase in a medium contains Na<sub>2</sub>HPO<sub>3</sub>•5H<sub>2</sub>0 (1.32mM) and NH<sub>4</sub>Cl serving as the phosphorus and nitrogen sources, respectively.</b>
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</center>
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The <i>E.coli</i> expressing phosphite dehydrogenase outcompeted a control strain in phosphite-containing media after 72 hours. However, the negative control group still grew in a certain degree. We looked up the researches and presumed that, alkaline phosphatase(BAP) found in <i>E.coli</i>, will be increasingly express when the amount of bio-available phosphorus is low. Through the hydrolysis, the BAP then turn the phosphorus compounds in the environment into bio-available phosphorus. With the help of the BAP, the microbes can utilize the phosphorus compounds more effectively, which also facilitates the modified pathways.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2325102 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2325102 SequenceAndFeatures</partinfo>
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===Usage and Biology===
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Using the pGEX-2T vector, the PTDH could be expressed in BL21(DE3).
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[[File:BBa_K2325102--1.png|thumb|left|400px|""Figure 1."" Growth curve of <i>E.coli</i> BL21(DE3) and the engineering <i>E.coli</i> BL21(DE3)(pGEX-ptx) expressing phosphite dehydrogenase in a medium contains Na<sub>2</sub>HPO<sub>3</sub>•5H<sub>2</sub>0 (1.32mM) and NH<sub>4</sub>Cl serving as the phosphorus and nitrogen sources, respectively.]]
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The <i>E.coli</i> expressing phosphite dehydrogenase outcompeted a control strain in phosphite-containing media after 72 hours. However, the negative control group still grew in a certain degree. We looked up the researches and presumed that, alkaline phosphatase(BAP) found in <i>E.coli</i>, will be increasingly express when the amount of bio-available phosphorus is low. Through the hydrolysis, the BAP then turn the phosphorus compounds in the environment into bio-available phosphorus. With the help of the BAP, the microbes can utilize the phosphorus compounds more effectively, which also facilitates the modified pathways.
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Latest revision as of 13:30, 29 October 2017


Phosphite dehydrogenase expression device

The BBa_K2325102 contains phosphite dehydrogenase gene with T7 promoter and terminator.

Phosphite dehydrogenase is an enzyme involved in the production of phosphate for providing phosphorus source for the engineering bacteria. This part has been constructed into expression vectors to modify the metabolic pathway of phosphorus in E.coli.


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]


Usage and Biology

Using the pGEX-2T vector, the PTDH could be expressed in BL21(DE3).

""Figure 1."" Growth curve of E.coli BL21(DE3) and the engineering E.coli BL21(DE3)(pGEX-ptx) expressing phosphite dehydrogenase in a medium contains Na2HPO3•5H20 (1.32mM) and NH4Cl serving as the phosphorus and nitrogen sources, respectively.

The E.coli expressing phosphite dehydrogenase outcompeted a control strain in phosphite-containing media after 72 hours. However, the negative control group still grew in a certain degree. We looked up the researches and presumed that, alkaline phosphatase(BAP) found in E.coli, will be increasingly express when the amount of bio-available phosphorus is low. Through the hydrolysis, the BAP then turn the phosphorus compounds in the environment into bio-available phosphorus. With the help of the BAP, the microbes can utilize the phosphorus compounds more effectively, which also facilitates the modified pathways.