Difference between revisions of "Part:BBa K5078001"

Revision as of 18:29, 28 September 2024

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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