Difference between revisions of "Part:BBa K2505001:Design"

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−	===Design Notes===	+	==Design Notes==
−	<span style="margin-left: 25px;">This part produces AHK4, a receptor of iP(isopentenyl adenine: kind of cytokinin). This part was introduced to <i>E.coli</i> KMI002 strain and sensed iP synthesized by human cell. The gene(<i>ahk4</i>) is derived from <i>A.thaliana</i> and optimized for <i>E.coli</i> codon in reference to the codon usage. The AHK4 protein is transmembrane protein and toxic for <i>E.coli</i> cell. Thus, We regulated expression of AHK4 by P<sub>BAD/araC</sub>: L-arabinose operon(tight transcriptionaly regulation system)	+	<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.
	===Source===		===Source===
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	===References===		===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.

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Latest revision as of 18:00, 1 November 2017

pBad/araC-rbs-ahk4

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.