Difference between revisions of "Part:BBa K4117888"

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

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4117888 short</partinfo>
-how to make 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([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.
 <!-- Add more about the biology of this part here