Difference between revisions of "Part:BBa K4883007"

 
 
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Insert a 2A self-cleaving peptide, PTV, between RIB7 and ADE4. ERBV-1 helps to achieve bicistronic gene expression in Saccharomyces cerevisiae.
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This is a riboflavin synthase RIB7, 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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===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. RIB7 ([https://parts.igem.org/Part:BBa_K4883002 BBa_K4883002]) catalyzes the second step of the riboflavin (vitamin B2) biosynthesis pathway in S. cerevisiae. Co-overexpression of RIB7 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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<center><img src = "https://static.igem.wiki/teams/4883/wiki/bba-k4883007-1.png" style = "width:380px"></center>
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<figcaption><center>Figure 1. Gel electrophoresis of PCR products of RIB7-PTV-ADE4.  </center></figcaption>
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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.
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Latest revision as of 07:26, 6 October 2023


RIB7-PTV-ADE4

This is a riboflavin synthase RIB7, 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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 240
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 172



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. RIB7 (BBa_K4883002) catalyzes the second step of the riboflavin (vitamin B2) biosynthesis pathway in S. cerevisiae. Co-overexpression of RIB7 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).

Figure 1. Gel electrophoresis of PCR products of RIB7-PTV-ADE4.

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.