Part:BBa_K5477035
UDPD-pGAL1/10-UGT2B15 detox module against BPA
This composite part consists of UDP-glucuronosyltransferase 2B15 (UGT2B15) BBa_K54770169 , pGAL1/10 bidirectional promoter BBa_K5477005 and UDP-glucose dehydrogenase BBa_K5477018. pGAL1/10 drives the expression of the two enzymes in the opposite direction. UDPD synthesizes the necessary precursor (UDP-glucoronic acid) that UGT2B15 uses for phase II detoxification of BPA (1) (2).
The 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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3466
Illegal PstI site found at 542 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3466
Illegal PstI site found at 542 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3466
Illegal BglII site found at 1122
Illegal BamHI site found at 3286
Illegal BamHI site found at 3580 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3466
Illegal PstI site found at 542 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3466
Illegal PstI site found at 542
Illegal AgeI site found at 1828 - 1000COMPATIBLE WITH RFC[1000]
References
1. Hanioka N, Naito T, Narimatsu S. Human UDP-glucuronosyltransferase isoforms involved in bisphenol A glucuronidation. Chemosphere. 2008 Dec 1;74(1):33–6.
2. Oka T, Jigami Y. Reconstruction of de novo pathway for synthesis of UDP-glucuronic acid and UDP-xylose from intrinsic UDP-glucose in Saccharomyces cerevisiae. FEBS J. 2006 Jun;273(12):2645–57.
| None |