Part:BBa_K4883003
RIB1-PTV-ADE4
This is a riboflavin synthase RIB1, connected with a PTV 2A self-cleaving peptide, followed by an IMP synthase ADE4.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 428
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 448
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
ADE4 (BBa_K4883000) catalyzes the first step of the purine nucleotide biosynthetic pathway in Saccharomyces cerevisiae, forming IMP. IMP is the precursor of GTP, and GTP is the precursor of vitamin B2. RIB1 (BBa_K4883001) catalyzes the first step of the riboflavin (vitamin B2) biosynthesis pathway. Co-overexpression of RIB1 and ADE4 could lead to overproduction of vitamin B2 in S. cerevisiae.
For the two genes to be expressed separately and normally, the genes are divided by a 2A self-cleaving peptide, PTV, which cleaves a longer peptide into two shorter peptides. PTV has been proven to have the second-highest cleavage efficiency in S. cerevisiae (Souza-Moreira et al., 2018).
References
Souza-Moreira TM, Navarrete C, Chen X, Zanelli CF, Valentini SR, Furlan M, Nielsen J, Krivoruchko A. Screening of 2A peptides for polycistronic gene expression in yeast. FEMS Yeast Res. 2018 Aug 1;18(5). doi: 10.1093/femsyr/foy036. PMID: 29617770.
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