Difference between revisions of "Part:BBa K5184019"

 
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In order to achieve enhanced effects, we aimed to produce 9HZ and 9H10epoZ also, which have better repellent effects than 7epiZ. Due to difficulties of expressing the oxidase and reductases in E. coli, we decide to change the chassis to S. cerevisiae. This part presents the collection of promoters, genes coding for the oxidase and reductase and terminator. This enzyme collection may provide future iGEM teams with an insight into the expression of an oxidase and a reductase in S. cerevisiae.
  
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==Abstract==
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===Usage and Biology===
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ShZPO is a cytochrome P450 oxygenase found in Solanum habrochaites. It carries out two successive oxidations to generate two sesquiterpenes from a monocyclic sesquiterpene as the susbtrate. SlCPR2 is a NADPH-cytochrome P450 reductase found in Solanum lycopersicum. SlCPR2 is co-localized with ShZPO on the ER membrane, ensuring efficient electron transfer. We used pTDH3 and tTDH3 for the expression of ShZPO; pPGK1 and tPGK1 for the expression of SlCPR2.
  
 
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<partinfo>BBa_K5184019 SequenceAndFeatures</partinfo>
 
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===Usage and Biology===
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ShZPO is a cytochrome P450 oxygenase found in Solanum habrochaites. It carries out two successive oxidations to generate two sesquiterpenes from a monocyclic sesquiterpene as the susbtrate. SlCPR2 is a NADPH-cytochrome P450 reductase found in Solanum lycopersicum. Consisting of two domains, one with a binding site for flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide phosphate (NADPH) and another with a binding site for flavin mononucleotide (FMN), SlCPR2 functions through transferring electrons to ShZPO.
  
 
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Revision as of 11:49, 30 September 2024


pTDH3-ShZPO-tTDH1-pPGK1-slCPR2-tPGK1

In order to achieve enhanced effects, we aimed to produce 9HZ and 9H10epoZ also, which have better repellent effects than 7epiZ. Due to difficulties of expressing the oxidase and reductases in E. coli, we decide to change the chassis to S. cerevisiae. This part presents the collection of promoters, genes coding for the oxidase and reductase and terminator. This enzyme collection may provide future iGEM teams with an insight into the expression of an oxidase and a reductase in S. cerevisiae.

Abstract

ShZPO is a cytochrome P450 oxygenase found in Solanum habrochaites. It carries out two successive oxidations to generate two sesquiterpenes from a monocyclic sesquiterpene as the susbtrate. SlCPR2 is a NADPH-cytochrome P450 reductase found in Solanum lycopersicum. SlCPR2 is co-localized with ShZPO on the ER membrane, ensuring efficient electron transfer. We used pTDH3 and tTDH3 for the expression of ShZPO; pPGK1 and tPGK1 for the expression of SlCPR2.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3525
    Illegal XhoI site found at 2177
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2423
    Illegal BsaI.rc site found at 3535

Usage and Biology

ShZPO is a cytochrome P450 oxygenase found in Solanum habrochaites. It carries out two successive oxidations to generate two sesquiterpenes from a monocyclic sesquiterpene as the susbtrate. SlCPR2 is a NADPH-cytochrome P450 reductase found in Solanum lycopersicum. Consisting of two domains, one with a binding site for flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide phosphate (NADPH) and another with a binding site for flavin mononucleotide (FMN), SlCPR2 functions through transferring electrons to ShZPO.