Difference between revisions of "Part:BBa K1036001:Design"

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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>Xba</i>I and the <i><b>ndh</b></i> digested by <i>Eco</i>R & <i>Spe</i>I were ligated together by T4 ligase.
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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.
 
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Latest revision as of 13:05, 27 September 2013


the ndh gene coding for respiratory NADH dehydrogenase II


Barcodes are discontinued, but one was appended to the sequence of this part. Composite parts using this part will include the barcode. More ...

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1360
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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.

Source

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

References

Messner, K. R. & Imlay, J. A. The identification of primary sites of superoxide and hydrogen peroxide formation in the aerobic respiratory chain and sulfite reductase complex of Escherichia coli. J. Biol. Chem. 274, 10119-10128 (1999)