Part:BBa_K1076001:Design
Flanking genes for FadR deletion
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 768
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
To design the upstream gene we used primers with Apa1(5') and NdeI(3') and to downsteam gene we used primers with NdeI(5') and BamH1(3')ends, this parts have Nde1 complementary site and they were ligate together. This method garantee generation of a deleted copy of the target gene using a two step assymmetric/crossover PCR amplification. The target in this case is the FadR.
Source
It comes from amplified genomic sequence
References
HEIDELBERG, John F. et al. Genome sequence of the dissimilatory metal ion–reducing bacterium Shewanella oneidensis. Nature biotechnology, v. 20, n. 11, p. 1118-1123, 2002.
GORBY, Yuri A. et al. Electrically conductive bacterial nanowires produced by Shewanella oneidensis strain MR-1 and other microorganisms.Proceedings of the National Academy of Sciences, v. 103, n. 30, p. 11358-11363, 2006.
SUKOVICH, David J. et al. Structure, function, and insights into the biosynthesis of a head-to-head hydrocarbon in Shewanella oneidensis strain MR-1. Applied and environmental microbiology, v. 76, n. 12, p. 3842-3849, 2010.
ROBINSON, Lee T. Fatty Acid Metabolic Engineering: Insights for Bacterial Hydrocarbon Production. 2012. Doctor tese UNIVERSITY OF MINNESOTA.
KAZAKOV, Alexey E. et al. Comparative genomics of regulation of fatty acid and branched-chain amino acid utilization in proteobacteria. Journal of bacteriology, v. 191, n. 1, p. 52-64, 2009.
EVANS, W. Charles; FUCHS, G. Anaerobic degradation of aromatic compounds. Annual Reviews in Microbiology, v. 42, n. 1, p. 289-317, 1988.