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Heterologous Expression of a Membrane‐Spanning Auxin Importer: Implications for Functional Analyses of Auxin Transporters
Author(s) -
David J. Carrier,
Norliza Tendot Abu Bakar,
Karen Lawler,
James Matthew Dorrian,
Ameena J. Haider,
Malcolm J. Bennett,
Ian D. Kerr
Publication year - 2009
Publication title -
international journal of plant genomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.454
H-Index - 30
eISSN - 1687-5370
pISSN - 1687-5389
DOI - 10.1155/2009/848145
Subject(s) - auxin , heterologous expression , transporter , heterologous , microbiology and biotechnology , chemistry , biology , biochemistry , gene , recombinant dna
Biochemical studies of plant auxin transporters in vivo are made difficult by the presence of multiple auxin transporters and auxin-interacting proteins. Furthermore, the expression level of most such transporters in plants is likely to be too low for purification and downstream functional analysis. Heterologous expression systems should address both of these issues. We have examined a number of such systems for their efficiency in expressing AUX1 from Arabidopsis thaliana . We find that a eukaryotic system based upon infection of insect cells with recombinant baculovirus provides a high level, easily scalable expression system capable of delivering a functional assay for AUX1. Furthermore, a transient transfection system in mammalian cells enables localization of AUX1 and AUX1-mediated transport of auxin to be investigated. In contrast, we were unable to utilise P. pastoris or L. lactis expression systems to reliably express AUX1.

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