Difference between revisions of "Part:BBa K4035015"

 
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This part is made of the Gal1 promoter (BBa_J63006), the A-agglutinin-binding subunit Aga2 (BBa_K416000) linked to the dimer protein CUP1-GGGGS(EAAAK)2GGGGS-CUP1 (BBa_K4035008) and the V5 tag (BBa_T2020).  
 
This part is made of the Gal1 promoter (BBa_J63006), the A-agglutinin-binding subunit Aga2 (BBa_K416000) linked to the dimer protein CUP1-GGGGS(EAAAK)2GGGGS-CUP1 (BBa_K4035008) and the V5 tag (BBa_T2020).  
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===Usage and Biology===
  
 
Copper metallotionein CUP1 (BBa_M45090) is a protein responsible for copper binding protein in the yeast Saccharomyces cerevisiae. In order to increase the copper retrieval efficiency, two copies of CUP1 were linked together and expressed at the outter surface of S. cerevisiae. Aga2 attaches to the yeast cell wall through disulfide bonds to Aga1p. The fusion of the dimer and Aga2 then leads to the presence of the CUP1 dimer on the outter membrane of the cell. This fusion protein also contained a V5 tag in order to check its expression by Western Blot and Immunostaining. The expression was under the control of the Gal1 promoter, so that the protein was expressed only in the presence of galactose.  
 
Copper metallotionein CUP1 (BBa_M45090) is a protein responsible for copper binding protein in the yeast Saccharomyces cerevisiae. In order to increase the copper retrieval efficiency, two copies of CUP1 were linked together and expressed at the outter surface of S. cerevisiae. Aga2 attaches to the yeast cell wall through disulfide bonds to Aga1p. The fusion of the dimer and Aga2 then leads to the presence of the CUP1 dimer on the outter membrane of the cell. This fusion protein also contained a V5 tag in order to check its expression by Western Blot and Immunostaining. The expression was under the control of the Gal1 promoter, so that the protein was expressed only in the presence of galactose.  
  
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===Characterization===
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===Usage and Biology===
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===Sequence and Features===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4035015 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4035015 SequenceAndFeatures</partinfo>
  

Revision as of 06:59, 13 September 2021


Expression of the CUP1-GGGGS(EAAAK)2GGGGS-CUP1 dimer at the outter membrane of S. cerevisiae

This part is made of the Gal1 promoter (BBa_J63006), the A-agglutinin-binding subunit Aga2 (BBa_K416000) linked to the dimer protein CUP1-GGGGS(EAAAK)2GGGGS-CUP1 (BBa_K4035008) and the V5 tag (BBa_T2020).

Usage and Biology

Copper metallotionein CUP1 (BBa_M45090) is a protein responsible for copper binding protein in the yeast Saccharomyces cerevisiae. In order to increase the copper retrieval efficiency, two copies of CUP1 were linked together and expressed at the outter surface of S. cerevisiae. Aga2 attaches to the yeast cell wall through disulfide bonds to Aga1p. The fusion of the dimer and Aga2 then leads to the presence of the CUP1 dimer on the outter membrane of the cell. This fusion protein also contained a V5 tag in order to check its expression by Western Blot and Immunostaining. The expression was under the control of the Gal1 promoter, so that the protein was expressed only in the presence of galactose.

Characterization

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1201