Part:BBa_K5327011:Design
UDP-glycosyltransferase 74B1
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 370
Illegal BamHI site found at 265 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The design of the UDP-glycosyltransferase 74B1 (UGT74B1) gene is based on the coding sequence (CDS) from Arabidopsis thaliana and has been codon-optimized for efficient expression in Saccharomyces cerevisiae (S288C). UGT74B1 plays a crucial role in glucosinolate biosynthesis, specifically participating in the glycosylation step. It is specific for the thiohydroximate functional group and does not glycosylate the carboxylate group or a hydroxyl group.[1]To ensure high-level expression and mRNA stability, the design employs the PGK1 promoter(BBa_K2637015) and ADH1 terminator(BBa_K2637012). The optimized gene will be cloned into a vector, introduced into yeast S288C through homologous recombination, and validated using a knockout strain. This design aims to enhance the production efficiency of glucosinolates in yeast and further optimize yeast as a platform for metabolic engineering.
Plasmid
- Fig 1. The plasmid expression of UDP-glycosyltransferase 74B1
Source
Arabidopsis thaliana
References
- ↑ GRUBB C D, ZIPP B J, LUDWIG-MüLLER J, et al. Arabidopsis glucosyltransferase UGT74B1 functions in glucosinolate biosynthesis and auxin homeostasis [J]. The Plant journal : for cell and molecular biology, 2004, 40(6): 893-908.