Part:BBa_K1356005:Design
Nitric oxide reductase (norB) from Pseudomonas aeruginosa PAO1
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1390
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 284
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 310
Illegal NgoMIV site found at 352
Illegal NgoMIV site found at 1254 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This gene was cloned from Pseudomonas aeruginosa PAO1 genomic DNA into pSB1C3 using the XbaI and SpeI restriction sites. Correct sequence and orientation were confirmed using 454 Pyrosequencing (BYU). The original sequence contained PstI sites starting at bases 115 and 1,231. These sequences were changed to CTTCAG and CTACAG, respectively, using site-specific mutagenesis; the mutant sites were verified to code for the same amino acids. Mutagenesis was also confirmed using 454 Pyrosequencing (BYU).
Source
This gene was cloned from Pseudomonas aeruginosa PAO1 genomic DNA, which was isolated from a bacterial stock provided by Dr. Stephen Lory at Harvard Medical School in Boston.
References
1. Z. Chen et al., Differentiated response of denitrifying communities to fertilization regime in paddy soil. Microbial ecology 63, 446 (Feb, 2012).
2. H. Arai, Regulation and Function of Versatile Aerobic and Anaerobic Respiratory Metabolism in Pseudomonas aeruginosa. Frontiers in microbiology 2, 103 (2011).
3. V. Kathiravan, Pseudomonas aeruginosa and Achromobacter sp.: nitrifying aerobic denitrifiers have a plasmid encoding for denitrifying functional genes. World journal of microbiology & biotechnology 30, 1187 (2014).