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dc.contributor.authorBlack, Moyra M.
dc.contributor.authorStockum, Christine
dc.contributor.authorDickson, James M.
dc.contributor.authorPutterill, Joanna
dc.contributor.authorArcus, Vickery L.
dc.coverage.spatialUnited Statesen_NZ
dc.date.accessioned2010-12-07T21:36:57Z
dc.date.available2010-12-07T21:36:57Z
dc.date.issued2010
dc.identifier.citationBlack, M.M., Stockum, C., Dickson, J.M., Putterill, J. & Arcus, V.L. (2010). Expression, purification and characterisation of GIGANTEA: A circadian clock-controlled regulator of photoperiodic flowering in plants. Protein Expression and Purification, available online 24 November 2010.en_NZ
dc.identifier.urihttps://hdl.handle.net/10289/4857
dc.description.abstractThe Arabidopsis thaliana (Arabidopsis) GIGANTEA (GI) gene is a central component of the photoperiodic flowering pathway. While it has been 40 years since the first mutant alleles of GI were described much is still unknown about the molecular mechanism of GI action. To investigate the biochemistry and domain organisation (and ultimately to give a greater understanding of the role of GI in floral induction), it is first necessary to produce significant quantities of purified protein. Soluble affinity-tagged full-length GI was expressed in Escherichia coli (E. coli) and was stabilised by the addition of the detergent n-dodecyl-β-d-maltoside (DDM) to storage and purification buffers. Stabilised GI was purified using a variety of chromatographic methods, and characterised using a selection of biochemical techniques including circular dichroism, and dynamic light scattering. This showed that purified GI contained secondary structure, but was polydisperse in solution. Electron microscopy suggests a possible tetramer arrangement of GI. Limited proteolytic digests and mass spectrometry were used to identify potential GI domains. This led to the identification of a predicted 46 kDa amino-terminal GI domain. GI was also expressed in Sf9 insect cells using the baculovirus expression system. GI produced via this route gave insoluble protein.en_NZ
dc.language.isoen
dc.publisherElsevieren_NZ
dc.subjectfloweringen_NZ
dc.subjectplant proteinsen_NZ
dc.subjectelectron microscopyen_NZ
dc.subjectcircular dichroismen_NZ
dc.subjectlow temperature expressionen_NZ
dc.titleExpression, purification and characterisation of GIGANTEA: A circadian clock-controlled regulator of photoperiodic flowering in plantsen_NZ
dc.typeJournal Articleen_NZ
dc.identifier.doi10.1016/j.pep.2010.11.009en_NZ
dc.relation.isPartOfProtein Expression and Purificationen_NZ
pubs.begin-page197en_NZ
pubs.elements-id36276
pubs.end-page204en_NZ
pubs.issue2en_NZ
pubs.volume76en_NZ


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