Difference between revisions of "Part:BBa K1036001:Design"
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===Design Notes=== | ===Design Notes=== | ||
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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. | |
| + | </body> | ||
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===Source=== | ===Source=== | ||
Revision as of 17:00, 26 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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1360
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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. The biobrick BBa_F2621(located in 2012-Plate 2-21F) digested by EcoR & XbaI and the ndh digested by EcoR & SpeI were ligated together by T4 ligase.
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)