Part:BBa_K5078001
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NosZ from Dechloromonas denitrificans
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Usage and Biology
NosZ-D.deniti is the Nitrous Oxide Reductase gene from Dechloromonas denitrificans , codon optimized for expression in Chlamydomonas reinhardtii. Nitrous oxide reductase is responsible for the reduction of nitrous oxide (N₂O) into water and dinitrogen (N₂) by acting as the catalysis of a copper-dependent two electron reduction of N₂O [1]. Which ensures that a bacterial host will not have a truncated nitrogen pathway, and will put harmless N₂ in the atmosphere instead of the greenhouse gas N₂O. Additionally this will case the bacterial host to uptake more nitrogen from its environment. Note NosZ-Denit is similar to NosZ-P.Stuzeri (BBa_K5078000).
Figure 1. NosZ-D.denti in a level 0 plasmid for later golden gate assembly.
Verification of Nosz-P.stu
Successful transformation of pL0-D.den into host bacterium can be determined by a restriction digestion with the restriction enzyme BbsI-HF, with expected band lengths being 2292bp and 2088bp. Additionally bacterial colonies should appear white in the present X-gal.
Figure 2. pL0-NosZ-D.denit diagnostic digest using BbsI on a 1% agarose gel. The restriction digest indicated that the two colonies taken from both culture 1 and 2 all have pL0-NosZ-D.denit.
Structure simulation
Figure 3. Structure prediction of nitrous oxide reductase by NosZ-D.denit using AlphaFold. The structure has two identical 65.8kDa subunits that each contain 6 copper atoms [1]. The dark blue is high confidence of the structure and red is low confidence.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 777
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 777
Illegal NotI site found at 2158 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1301
Illegal BamHI site found at 227
Illegal BamHI site found at 965
Illegal XhoI site found at 580
Illegal XhoI site found at 697 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 777
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 777
Illegal AgeI site found at 160
Illegal AgeI site found at 1403 - 1000COMPATIBLE WITH RFC[1000]
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