Difference between revisions of "Part:BBa K2505001:Design"
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===Design Notes===
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==Design Notes==
 
<p>This gene is derived from <i>Arabidopsis thaliana</i> and encode a receptor of cytokinins, AHK4. Cytokinins are signaling molecules (Phytohormones)  in plants and play important roles in cell growth and differentiation.  AHK4 has a histidine kinase activity, and binding of a cytokinin to AHK4 triggers auto-phosphorylation of AHK4 and the following histidine-to-aspartate phosphorelay. As a consequence, transcription from target genes is induced and/or repressed so that physiological states of plants are changed. Since overexpression of AHK4 seemes to be toxic for <i>E. coli</i>, the expression is tightly regulated by BAD/<i>araC</i> promoter, an L-arabinose inducible promoter.</p>
 
<p>This gene is derived from <i>Arabidopsis thaliana</i> and encode a receptor of cytokinins, AHK4. Cytokinins are signaling molecules (Phytohormones)  in plants and play important roles in cell growth and differentiation.  AHK4 has a histidine kinase activity, and binding of a cytokinin to AHK4 triggers auto-phosphorylation of AHK4 and the following histidine-to-aspartate phosphorelay. As a consequence, transcription from target genes is induced and/or repressed so that physiological states of plants are changed. Since overexpression of AHK4 seemes to be toxic for <i>E. coli</i>, the expression is tightly regulated by BAD/<i>araC</i> promoter, an L-arabinose inducible promoter.</p>
 
<p>The DNA sequences of this gene is optimized for expressing in <i>E. coli</i> cells considering the codon usage.
 
<p>The DNA sequences of this gene is optimized for expressing in <i>E. coli</i> cells considering the codon usage.

Revision as of 18:00, 31 October 2017


pBad/araC-rbs-ahk4


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1294
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2375
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961
    Illegal SapI site found at 1542
    Illegal SapI.rc site found at 3177


Design Notes

This gene is derived from Arabidopsis thaliana and encode a receptor of cytokinins, AHK4. Cytokinins are signaling molecules (Phytohormones) in plants and play important roles in cell growth and differentiation. AHK4 has a histidine kinase activity, and binding of a cytokinin to AHK4 triggers auto-phosphorylation of AHK4 and the following histidine-to-aspartate phosphorelay. As a consequence, transcription from target genes is induced and/or repressed so that physiological states of plants are changed. Since overexpression of AHK4 seemes to be toxic for E. coli, the expression is tightly regulated by BAD/araC promoter, an L-arabinose inducible promoter.

The DNA sequences of this gene is optimized for expressing in E. coli cells considering the codon usage.

Source

This subparts were synthesized by IDT or used from iGEM Desrtibutuion kits.

References

Suzuki, T., Miwa, K., Ishikawa, K., Yamada, H., Aiba, H. and Mizuno, T. (2001) The Arabidopsis Sensor His-kinase, AHK4, Can Respond to Cytokinins. Plant Cell Physiol. 42: 107-113.

Yamada, H., Suzuki, T., Terada, K., Takei, K., Ishikawa, K., Miwa, K., Yamashino, T. and Mizuno, T. (2001) The Arabidopsis AHK4 Histidine Kinases is a Cytokinin-Binding Receptor that Transduces Cytokinin Signals Across the Membrane. Plant Cell Physiol. 42: 1017-1023.

Spíchal, L., Rakova, N.Y., Riefler, M., Mizuno, T., Romanov, G.A.,Strnad, M. and Schmülling, T. (2004) Two Cytokinin Receptors of Arabidopsis thaliana, CRE1/AHK4 and AHK3, Differ in their Ligand Specifity in a Bacterial Assay. Plant Cell Physiol. 45: 1299-1305.

Klimeš, P., Turek, D., Mazura, P., Gallová, L., Spíchal, L. and Brzobohatý, B. (2017) High Throughput Screening Method for Identifying Potential Agonists and Antagonists of Arabidopsis thaliana Cytokinin Receptor CRE1/AHK4. Frontiers in Plant Science.

Mizuno, T. and Yamashino, T. (2010) BIOCHEMICAL CHARACTERIZATION OF PLANT HORMONE CYTOKININ-RECEPTOR HISTIDINE KINASES USING MICROORGANISMS. Methods in Enzymology: 335-344.

Nakashima, N., Akita, H. and Hoshino, T. (2014) Establishment of a novel gene expression method, BICES (biomass-inducible chromosome-based expression system), and its application to the production of 2,3-butanediol and acetoin. Metab Eng. 25 :204-214.