Part:BBa_K5078000

Nos Z gene from P. stutzeri

This is the Nitrous Oxide Reductase gene from P. stutzeri codon optimized for expression in Chlamydomonas reinhardtii

Figure 1. NosZ P.stutzeri in a level 0 plasmid for later golden gate assembly.

Verification of nosZ P. stu

Successful transformation of pL0-P.stu into host bacterium can be determined by a restriction digestion with the restriction enzyme BbsI, with the molecular weights being 1914bp and 2088bp. Additionally bacterial colonies should appear white in the present X-gal.

Figure 2. pL0-NosZ-P.stuzeri diagnostic digest using BbsI on a 8% agarose gel. The restriction digest indicated that the two colonies taken from both culture 1 and 2 all have pL0-NosZ-P.stuzeri.

Structure simulation

Figure 3. Structure prediction of nitrous oxide reductase. The structure has two identical 65.8kDa subunits that each contain 6 copper atoms

References

[1] Wan, S., Johnson, A. M., & Altosaar, I. (2012). Expression of nitrous oxide reductase from Pseudomonas stutzeri in transgenic tobacco roots using the root-specific rolD promoter from Agrobacterium rhizogenes. Ecology and evolution, 2(2), 286–297. https://doi.org/10.1002/ece3.74 Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 541
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1303
  Illegal PstI site found at 541
  Illegal NotI site found at 145
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1520
  Illegal BamHI site found at 467
  Illegal XhoI site found at 34
  Illegal XhoI site found at 1588
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 541
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 541
  Illegal NgoMIV site found at 1075
  
• 1000
  COMPATIBLE WITH RFC[1000]