Difference between revisions of "Part:BBa K3091000"

 
 
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===Usage and Biology===
 
===Usage and Biology===
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<h1>Aim</h1>
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<p>We built this new part as an approach to detecting bacteriophages in fecal polluted water through color change. This enzyme is from Solanum tuberosum, and would cause yellow solution containing tea polyphenol turn red. Also, the product of the reaction, theaflavins, has multiple health benefits. Our bacteria can potentially also play a part in theaflavins industrial production.</p>
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<h1>Plasmid Construction</h1>
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<p>We acquired the gene sequence through NCBI blast, and synthesized the sequence thanks to Twist, and selected pET expression system due to its high efficiency and capability to be regulated by IPTG induction.</p>
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<p>We tried to use digestion and ligation to construct the plasmid, but it failed numerous times. We speculate the reason is that the efficiency of the enzyme Bgl2 is lowered due to long time storage in the lab. Then we borrowed in-fusion mix from BNU and built the plasmid again, and finally succeed.</p>
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<h1>Measurement and Reaction</h1>
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<p>1. Model</p>
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<p>We have documented the OD values when conducting IPTG induction to get the best level of protein expression. As induction proceeds and OD600 value reaches somewhat near 0.3, protein expression is tested to be at high level by mathematical model.</p>
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<p>2. Solution and Reaction</p>
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<p>Through various journals, we learned that potato PPO is most active at PH=5.5, temperature about 40~50°C. As for the substrate concentration, we still need to conduct control experiments because tea polyphenol can potentially have anti-bacteria effects. More tests should be done to find an ideal solution that make the reaction visible and reliable.</p>
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Latest revision as of 03:47, 22 October 2019


Polyphenol Oxidase [PPO]

Polyphenol oxidase is the biocatalyst of tea polyphenol and is able to catalyze redox reactions to form quinone as final product. The reaction is chromogenic and is widely observed in plants such as apple, litchi, potato and spinach. Part BBa_K3091000's sequence comes from PPO found in potato.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1180
    Illegal PstI site found at 434
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1180
    Illegal PstI site found at 434
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1180
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1180
    Illegal PstI site found at 434
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1180
    Illegal PstI site found at 434
    Illegal AgeI site found at 1009
  • 1000
    COMPATIBLE WITH RFC[1000]