Difference between revisions of "Part:BBa K4883003"

 
 
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Insert a 2A self-cleaving peptide, PTV, between RIB1 and ADE4. PTV helps to achieve bicistronic gene expression in Saccharomyces cerevisiae.
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This is a riboflavin synthase RIB1, connected with a PTV 2A self-cleaving peptide, followed by an IMP synthase ADE4.  
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===Usage and Biology===
 
  
 
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<partinfo>BBa_K4883003 parameters</partinfo>
 
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===Usage and Biology===
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ADE4 ([https://parts.igem.org/Part:BBa_K4883000 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 ([https://parts.igem.org/Part:BBa_K4883001 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.
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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).
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==References==
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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.

Latest revision as of 01:51, 6 October 2023


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 428
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 448
  • 1000
    COMPATIBLE 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.