Difference between revisions of "Part:BBa K1640020:Design"

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===Design Notes===
 
===Design Notes===
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Modularisation: This part is to be combined with psbD, containing psbD, and psbC, containing psbC, to form psbDCA, one of five operons for the biosynthesis of a minimal Photosystem II.
 
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Overview: When induced, Escherichia coli will express the D1 subunit of the photosystem II reaction centre.
 
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Detection Methods: PsbMZH has been detected in E. coli by gel electrophoresis, and confirmed by DNA sequencing.
 
  
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===Source===
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Genes required for photosystem II were selected using <html><a href=http://www.biocyc.org/CHLAMY/NEW-IMAGE?type=ENZYME&object=CPLX4LZ-235>biocyc entry on <i>Chlamydomonas reinhardtii</i> photosystem II complex</a>. Sequences were obtained from NCBI, targeting sequences were identified in nuclear using <a href=http://www.cbs.dtu.dk/services/ChloroP>ChloroP</a>, and removed, with an ATG added. Resulting sequences were codon optimized for e. coli expression using <a href=http://genomes.urv.es/OPTIMIZER/>Optimizer</a></html>, avoiding EcoRI, XbaI, SpeI and XbaI sites.
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''Chlamydomonas reinhardtii''
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===References===
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Ribosome binding sites were added in front of each CDS, and a promoter was added to biobricks which form the first part of their intended operon.
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Chu, H.-A., Debus, R. J., & Babcock, G. T. (2001). D1-Asp170 Is Structurally Coupled to the Oxygen Evolving Complex in Photosystem II As Revealed by Light-Induced Fourier Transform Infrared Difference Spectroscopy†. Biochemistry, 40(7), 2312-2316. doi:10.1021/bi0022994
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Erickson, J. M., Pfister, K., Rahire, M., Togasaki, R. K., Mets, L., & Rochaix, J. D. (1989). Molecular and biophysical analysis of herbicide-resistant mutants of Chlamydomonas reinhardtii: structure-function relationship of the photosystem II D1 polypeptide. The Plant Cell, 1(3), 361-371.
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Ferreira, K. N., Iverson, T. M., Maghlaoui, K., Barber, J., & Iwata, S. (2004). Architecture of the photosynthetic oxygen-evolving center. Science, 303(5665), 1831-1838.
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Jansen, M. A. K., Mattoo, A. K., Malkin, S., & Edelman, M. (1993). Direct Demonstration of Binding-Site Competition between Photosystem II Inhibitors at the QB Niche of the D1 Protein. Pesticide Biochemistry and Physiology, 46(1), 78-83. doi:http://dx.doi.org/10.1006/pest.1993.1039
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Marder, J., Chapman, D., Telfer, A., Nixon, P., & Barber, J. (1987). Identification of psbA and psbD gene products, D1 and D2, as reaction centre proteins of photosystem 2. Plant molecular biology, 9(4), 325-333.
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Nixon, P. J., & Diner, B. A. (1992). Aspartate 170 of the photosystem II reaction center polypeptide D1 is involved in the assembly of the oxygen-evolving manganese cluster. Biochemistry, 31(3), 942-948. doi:10.1021/bi00118a041
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Latest revision as of 08:26, 18 September 2015

psbA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Genes required for photosystem II were selected using biocyc entry on Chlamydomonas reinhardtii photosystem II complex. Sequences were obtained from NCBI, targeting sequences were identified in nuclear using ChloroP, and removed, with an ATG added. Resulting sequences were codon optimized for e. coli expression using Optimizer, avoiding EcoRI, XbaI, SpeI and XbaI sites.

Ribosome binding sites were added in front of each CDS, and a promoter was added to biobricks which form the first part of their intended operon.