Difference between revisions of "Part:BBa K4883011"

 
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This is a complete expression cassette consisting of a strong promoter Ptef1, an ADE4 gene, and a CYC1 terminator.  
 
This is a complete expression cassette consisting of a strong promoter Ptef1, an ADE4 gene, and a CYC1 terminator.  
  
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===Usage and Biology===
 
  
 
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<partinfo>BBa_K4883011 parameters</partinfo>
 
<partinfo>BBa_K4883011 parameters</partinfo>
 
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<!-- Add more about the biology of this part here -->
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===Usage and Biology===
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To mitigate vitamin B2 deficiency, we planned to make vitamin B2-enriched bread with engineered yeast, Saccharomyces cerevisiae. Overexpression of ADE4 (BBa_K4883000) should lead to the overproduction of vitamin B2 in S. cerevisiae. Ptef1 (BBa_K2407300) has been proven one of the strongest promoters of S. cerevisiae, so we used it for overexpression (Partow et al., 2010). A CYC1 terminator was also included to form the complete expression cassette. 
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===Characterization===
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'''2023 Hangzhou-SDG Team characterized this part with vitamin B2 production'''
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''Tests in Liquid YPD Media''
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The WT S. cerevisiae S288C and the strain overexpressing ADE4 were inoculated into YPD media, and cultured at 30 ℃, 180 rpm. The riboflavin concentrations in the supernatants were measured using high-performance liquid chromatography (HPLC).

Revision as of 02:05, 6 October 2023


Ptef1-ADE4-Tcyc1

This is a complete expression cassette consisting of a strong promoter Ptef1, an ADE4 gene, and a CYC1 terminator.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2197
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 176



Usage and Biology

To mitigate vitamin B2 deficiency, we planned to make vitamin B2-enriched bread with engineered yeast, Saccharomyces cerevisiae. Overexpression of ADE4 (BBa_K4883000) should lead to the overproduction of vitamin B2 in S. cerevisiae. Ptef1 (BBa_K2407300) has been proven one of the strongest promoters of S. cerevisiae, so we used it for overexpression (Partow et al., 2010). A CYC1 terminator was also included to form the complete expression cassette.


Characterization

2023 Hangzhou-SDG Team characterized this part with vitamin B2 production

Tests in Liquid YPD Media The WT S. cerevisiae S288C and the strain overexpressing ADE4 were inoculated into YPD media, and cultured at 30 ℃, 180 rpm. The riboflavin concentrations in the supernatants were measured using high-performance liquid chromatography (HPLC).