Difference between revisions of "Part:BBa K4117888"
 
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“T7 promoter+ pmrA+ RBS+ pmrB+ rrnB T1 terminator+ pmrC promoter” is the THC sensing pathway. To alert vaccinators that they have ingested Δ9-THC, we design a Δ9-THC sensing system. In this pathway, anti-THC can specifically bind to Δ9-THC, and then promote PmrB phosphorylation of PmrA. PmrC promoters activate phosphorylated PmrA proteins and initiate transcription to express UGT1A3.
 
“T7 promoter+ pmrA+ RBS+ pmrB+ rrnB T1 terminator+ pmrC promoter” is the THC sensing pathway. To alert vaccinators that they have ingested Δ9-THC, we design a Δ9-THC sensing system. In this pathway, anti-THC can specifically bind to Δ9-THC, and then promote PmrB phosphorylation of PmrA. PmrC promoters activate phosphorylated PmrA proteins and initiate transcription to express UGT1A3.
  
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Outer membrane protease T (ompT) is a 33.5 kDa endoprotease located on the outer membrane of Escherichia coli. OmpT signal peptide is an excretion tag linked on the N-terminus of protein. It can transport proteins across the outer membrane with the help of ompT.
+
Outer membrane protease T (ompT) is a 33.5 kDa endoprotease located on the outer membrane of ''Escherichia coli''. OmpT signal peptide is an excretion tag linked on the N-terminus of protein. It can transport proteins across the outer membrane with the help of ompT.
  
 
UGT1A3 in this pathway plays a significant role in metabolic section, working upon the sensing section. Tetrahydrocannabinol (Δ9 -THC), the primary psychoactive ingredient in marijuana, is subject to cytochrome P450 oxidation and subsequent UDP-glucuronosyltransferase(UGT)-dependent glucuronidation. Many studies have shown that CYP2C9 and CYP2C19 are the primary enzymes responsible for these cytochrome P450-dependent oxidations. In Mazur’s study, the specific human UGTs are responsible for classic cannabinoid metabolism, and among them, the highest activity toward THC-COOH was observed with UGT1A3.
 
UGT1A3 in this pathway plays a significant role in metabolic section, working upon the sensing section. Tetrahydrocannabinol (Δ9 -THC), the primary psychoactive ingredient in marijuana, is subject to cytochrome P450 oxidation and subsequent UDP-glucuronosyltransferase(UGT)-dependent glucuronidation. Many studies have shown that CYP2C9 and CYP2C19 are the primary enzymes responsible for these cytochrome P450-dependent oxidations. In Mazur’s study, the specific human UGTs are responsible for classic cannabinoid metabolism, and among them, the highest activity toward THC-COOH was observed with UGT1A3.
  
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Thus, in our system, Δ-9-THC is hydroxylated by CYP2C9(BBa_K4117002) and CYP2C19(BBa_K4117022), remaining psychoactive. The major product of CYP2C9 and CYP2C19 metabolism, 11-OH-Δ-9-THC, was further oxidized to THC-COOH. Then UGT1A3, which catalyzes the glucuronidation of the former substrate, increases the metabolite's water solubility, thereby facilitating excretion into the urine.
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Thus, in our system, Δ-9-THC is hydroxylated by CYP2C9([https://parts.igem.org/Part:BBa_K4117002, BBa_K4117002]) and CYP2C19([https://parts.igem.org/Part:BBa_K4117022, BBa_K4117022]), remaining psychoactive. The major product of CYP2C9 and CYP2C19 metabolism, 11-OH-Δ-9-THC, was further oxidized to THC-COOH. Then UGT1A3, which catalyzes the glucuronidation of the former substrate, increases the metabolite's water solubility, thereby facilitating excretion into the urine.
  
 
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Latest revision as of 08:32, 12 October 2022


UGT1A3

“T7 promoter+ pmrA+ RBS+ pmrB+ rrnB T1 terminator+ pmrC promoter” is the THC sensing pathway. To alert vaccinators that they have ingested Δ9-THC, we design a Δ9-THC sensing system. In this pathway, anti-THC can specifically bind to Δ9-THC, and then promote PmrB phosphorylation of PmrA. PmrC promoters activate phosphorylated PmrA proteins and initiate transcription to express UGT1A3.

Outer membrane protease T (ompT) is a 33.5 kDa endoprotease located on the outer membrane of Escherichia coli. OmpT signal peptide is an excretion tag linked on the N-terminus of protein. It can transport proteins across the outer membrane with the help of ompT.

UGT1A3 in this pathway plays a significant role in metabolic section, working upon the sensing section. Tetrahydrocannabinol (Δ9 -THC), the primary psychoactive ingredient in marijuana, is subject to cytochrome P450 oxidation and subsequent UDP-glucuronosyltransferase(UGT)-dependent glucuronidation. Many studies have shown that CYP2C9 and CYP2C19 are the primary enzymes responsible for these cytochrome P450-dependent oxidations. In Mazur’s study, the specific human UGTs are responsible for classic cannabinoid metabolism, and among them, the highest activity toward THC-COOH was observed with UGT1A3.

Thus, in our system, Δ-9-THC is hydroxylated by CYP2C9(BBa_K4117002) and CYP2C19(BBa_K4117022), remaining psychoactive. The major product of CYP2C9 and CYP2C19 metabolism, 11-OH-Δ-9-THC, was further oxidized to THC-COOH. Then UGT1A3, which catalyzes the glucuronidation of the former substrate, increases the metabolite's water solubility, thereby facilitating excretion into the urine.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4599
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4599
    Illegal BglII site found at 4257
    Illegal BamHI site found at 2887
    Illegal BamHI site found at 3444
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
    Illegal AgeI site found at 3031
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 732