Difference between revisions of "Part:BBa K5477038"

Revision as of 02:11, 2 October 2024

CYP3A4-pGAL1/10-POR detox module against a wide array of contaminants

The CYP3A4-pGAL1/10-POR composite part consists the CYP3A4 enzyme BBa_K5477020 and cytochrome P450 oxidoreductase (POR) BBa_K5477022 to form a detoxification module. The pGAL1/10 bidirectional promoter drives the co-expression of CYP3A4 and POR in opposite directions. CYP3A4, a phase I enzyme, is responsible for oxidizing around 50% of all clinically used drugs (1) (2) (3) (4). POR provides the electrons required for these oxidation reactions by transferring electrons from NADPH to CYP3A4 (5).

This composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 1483
Illegal PstI site found at 2486
Illegal PstI site found at 2683
Illegal PstI site found at 3067
Illegal PstI site found at 3507
Illegal PstI site found at 3567
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 2486
Illegal PstI site found at 2683
Illegal PstI site found at 3067
Illegal PstI site found at 3507
Illegal PstI site found at 3567
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 1483
Illegal PstI site found at 2486
Illegal PstI site found at 2683
Illegal PstI site found at 3067
Illegal PstI site found at 3507
Illegal PstI site found at 3567
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 1483
Illegal PstI site found at 2486
Illegal PstI site found at 2683
Illegal PstI site found at 3067
Illegal PstI site found at 3507
Illegal PstI site found at 3567
Illegal NgoMIV site found at 2889
Illegal NgoMIV site found at 3008
Illegal AgeI site found at 1894
1000
COMPATIBLE WITH RFC[1000]

References

1. Bardal SK, Waechter JE, Martin DS. Chapter 6 - Pharmacogenetics. In: Bardal SK, Waechter JE, Martin DS, editors. Applied Pharmacology [Internet]. Philadelphia: W.B. Saunders; 2011. p. 53–8. Available from: https://www.sciencedirect.com/science/article/pii/B9781437703108000063

2. Guengerich, F.. (2008). Cytochrome P450 and Chemical Toxicology. Chemical research in toxicology. 21. 70-83. 10.1021/tx700079z.

3. Klyushova LS, Perepechaeva ML, Grishanova AY. The Role of CYP3A in Health and Disease. Biomedicines. 2022 Oct 24;10(11):2686. doi: 10.3390/biomedicines10112686. PMID: 36359206; PMCID: PMC9687714.

4. Lynch T, Price A. The effect of cytochrome P450 metabolism on drug response, interactions, and adverse effects. Am Fam Physician. 2007;76(3):391-396.

5. Pandey AV, Flück CE. NADPH P450 oxidoreductase: structure, function, and pathology of diseases. Pharmacol Ther. 2013;138(2):229-254. doi:10.1016/j.pharmthera.2013.01.010

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K5477038 short</partinfo>
-In our system, the CYP3A4-pGAL1/10-POR composite part combines the CYP3A4 enzyme and cytochrome P450 oxidoreductase (POR) to form a highly efficient detoxification unit. The pGAL1/10 bidirectional promoter drives the expression of both CYP3A4 and POR simultaneously but in opposite directions. CYP3A4, a key phase I enzyme, is responsible for oxidizing around 50% of all clinically used drugs and environmental toxins. POR provides the electrons required for these oxidation reactions by transferring electrons from NADPH to CYP3A4, ensuring efficient metabolism of harmful xenobiotics and pharmaceuticals.
+The CYP3A4-pGAL1/10-POR composite part consists the CYP3A4 enzyme [https://parts.igem.org/Part:BBa_K5477020 BBa_K5477020] and cytochrome P450 oxidoreductase (POR) [https://parts.igem.org/Part:BBa_K5477022 BBa_K5477022] to form a detoxification module. The pGAL1/10 bidirectional promoter drives the co-expression of CYP3A4 and POR in opposite directions. CYP3A4, a phase I enzyme, is responsible for oxidizing around 50% of all clinically used drugs (1) (2) (3) (4). POR provides the electrons required for these oxidation reactions by transferring electrons from NADPH to CYP3A4 (5).
 This composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid.
@@ Line 21: / Line 21: @@
 <partinfo>BBa_K5477038 parameters</partinfo>
 <!-- -->
+===References===
+. Bardal SK, Waechter JE, Martin DS. Chapter 6 - Pharmacogenetics. In: Bardal SK, Waechter JE, Martin DS, editors. Applied Pharmacology [Internet]. Philadelphia: W.B. Saunders; 2011. p. 53–8. Available from: https://www.sciencedirect.com/science/article/pii/B9781437703108000063
+. Guengerich, F.. (2008). Cytochrome P450 and Chemical Toxicology. Chemical research in toxicology. 21. 70-83. 10.1021/tx700079z.
+. Klyushova LS, Perepechaeva ML, Grishanova AY. The Role of CYP3A in Health and Disease. Biomedicines. 2022 Oct 24;10(11):2686. doi: 10.3390/biomedicines10112686. PMID: 36359206; PMCID: PMC9687714.
+. Lynch T, Price A. The effect of cytochrome P450 metabolism on drug response, interactions, and adverse effects. Am Fam Physician. 2007;76(3):391-396.
+. Pandey AV, Flück CE. NADPH P450 oxidoreductase: structure, function, and pathology of diseases. Pharmacol Ther. 2013;138(2):229-254. doi:10.1016/j.pharmthera.2013.01.010