Difference between revisions of "Part:BBa K5078000"

(Structure simulation)
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<img src="https://static.igem.wiki/teams/5078/modeling/p-stu-nosz-folding-sphere.webp" width="400" height="auto"/><br>
 
<img src="https://static.igem.wiki/teams/5078/modeling/p-stu-nosz-folding-sphere.webp" width="400" height="auto"/><br>
 
Figure 3. Structure prediction of nitrous oxide reductase. The structure has two identical 65.8kDa subunits that each contain 6 copper atoms</div></html>
 
Figure 3. Structure prediction of nitrous oxide reductase. The structure has two identical 65.8kDa subunits that each contain 6 copper atoms</div></html>
 
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====References====
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[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
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 01:05, 22 September 2024


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]