Difference between revisions of "Part:BBa K1036000:Design"

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The <i>ndh</i> gene was amplifed from the genome of <i>E. coli</i> strain BL21(DE3) via PCR. The  
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The <i>ndh</i> gene was amplifed from the genome DNA of <i>E. coli</i> strain BL21(DE3) via PCR. Then RBS(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_B0034">BBa_B0034</a>), LVA-tag and terminator(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_B0015">BBa_B0015</a>) were fused with <i>ndh</i> via fusion PCR becoming RBS-<i>ndh</i>-LVA-TT marked as <i><b>ndh</b></i>. The DH5α colony carried <i>ndh</i> gene was sequenced after TA-cloning. The biobrick <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_F2621">BBa_F2621</a>(located in 2012-Plate 2-21F) digested by <i>Eco</i>R I & <i>Xba</i>I and the <i><b>ndh</b></i> digested by <i>Eco</i>R I & <i>Spe</i>I were ligated together by T4 ligase.
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And we know that the biobrick BBa_F2621 (position in plate: 2012-Plate 2-21F) is a quorum sensing promoter. We digested ''ndh'' with ''EcoR''I and ''Xba'' I, and 21F with ''EcoR''I and ''Spe''I. Then we constructed ''ndh'' plasmid by ligation.
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===References===
 
===References===
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1. Prindle, A., Samayoa, P., Razinkov, I., Danino, T., Tsimring, L.S., & Hasty, J. A sensing array of radically coupled genetic 'biopixels'. Nature 481, 39−44. (2012)
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<a href="http://www.ncbi.nlm.nih.gov/pubmed?term=A%20sensing%20array%20of%20radically%20coupled%20genetic%20'biopixels'.[all]&cmd=correctspelling">Prindle, A., Samayoa, P., Razinkov, I., Danino, T., Tsimring, L.S., & Hasty, J. A sensing array of radically coupled genetic 'biopixels'. Nature 481, 39−44. (2012)</a>
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Latest revision as of 21:43, 27 September 2013


lux pL controlled luxR with lux pR controlled ndh (LVA-tag) coding for NADH dehydrogenase II


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2544
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1101


Design Notes

The ndh gene was amplifed from the genome DNA of E. coli strain BL21(DE3) via PCR. Then RBS(BBa_B0034), LVA-tag and terminator(BBa_B0015) were fused with ndh via fusion PCR becoming RBS-ndh-LVA-TT marked as ndh. The DH5α colony carried ndh gene was sequenced after TA-cloning. The biobrick BBa_F2621(located in 2012-Plate 2-21F) digested by EcoR I & XbaI and the ndh digested by EcoR I & SpeI were ligated together by T4 ligase.

Source

The ndh gene was amplifed from the genome DNA of E. coli strain BL21(DE3) via PCR.

References

Prindle, A., Samayoa, P., Razinkov, I., Danino, T., Tsimring, L.S., & Hasty, J. A sensing array of radically coupled genetic 'biopixels'. Nature 481, 39−44. (2012)