Part:BBa_K1036001

the ndh gene coding for respiratory NADH dehydrogenase II

Description

ndh is the coding gene for NADH dehydrogenase II (NCBI), a non-proton-translating single subunit membrane bond enzyme important in maintaining a redox-balanced dinucleotide pool. NADH dehydrogenase II is the preponderant respiratory source of H₂O₂ as well as O₂^-. In fact, substantial H₂O₂ was formed when quinoneless vesicles containing only NADH dehydrogenase II were incubated with NADH.
Earlier studies estimated that the formation rate of intracellular O₂^- was about 10^-5 M S^-1 by extrapolation of O₂^- yield from this enzyme. And NADH dehydrogenase II formed about 4 O₂^- per 10 molecules of H₂O₂ both during respiration and in the quinoneless membranes.
In our oscillatoral circuit, ndh just like a "conductor" because this gene could express NDH-2 which is a membrane-bound respiratory enzyme that produces low levels of H₂O₂ and superoxide. H₂O₂ will trigger synchronized oscillation between colonies.

Experimental Data

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.
Construciton

Sequence and Features

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]