Difference between revisions of "Part:BBa K1189037:Design"
 
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<h1>Source</h1>
 
<h1>Source</h1>
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<p>We synthesized heavy ferrtin <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189019">(BBa_K1189019)</a> and light ferritin <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189020">(BBa_K1189020)</a> from a commerical synthesis company. These parts were optimized for expression in <i>E. coli K12</i>. These two elements were extracted via PCR, the pSB1C3 backbone was isolated with PCR, and these linear fragments were assembled into BBa_K1189037 using isothermal assembly.</p>
  
 
<h1>References</h1>
 
<h1>References</h1>
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<li>Chasteen, N. D., & Harrison, P. M. (1999). Mineralization in ferritin: an efficient means of iron storage. Journal of structural biology, 126(3), 182-194.</li>
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<li>Dehal, P. K., Livingston, C. F., Dunn, C. G., Buick, R., Luxton, R., & Pritchard, D. J. (2010). Magnetizable antibody‐like proteins. Biotechnology journal, 5(6), 596-604.</li>
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Latest revision as of 23:38, 31 October 2013

His tag + Human ferritin di-subunit with E coil under LacI promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 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 1307


Design notes

This part was created by fusing the heavy chain (BBa_K1189025, P02794 [UniParc]) and light chains (BBa_K1189024, P02792 [UniParc]) of human ferritin together. It is expressed under the LacI promoter (BBa_J04500) and has a his-tag for protein purification. An E-coil (BBa_K1189011) is included in order to allow binding of parts containing the respective K-coil (BBa_K1189010). This particular part is expressed under a strong RBS and the LacI promoter. The iGEM Calgary 2013 team attempted expression and purification of E coil ferritin di-subunit using this system. Additionally, they tested this same coding sequence under the T7 promoter in a different vector.

Source

We synthesized heavy ferrtin (BBa_K1189019) and light ferritin (BBa_K1189020) from a commerical synthesis company. These parts were optimized for expression in E. coli K12. These two elements were extracted via PCR, the pSB1C3 backbone was isolated with PCR, and these linear fragments were assembled into BBa_K1189037 using isothermal assembly.

References

  • Chasteen, N. D., & Harrison, P. M. (1999). Mineralization in ferritin: an efficient means of iron storage. Journal of structural biology, 126(3), 182-194.
  • Dehal, P. K., Livingston, C. F., Dunn, C. G., Buick, R., Luxton, R., & Pritchard, D. J. (2010). Magnetizable antibody‐like proteins. Biotechnology journal, 5(6), 596-604.