Overexpression of Brassinosteroid Regulators

The cDNAs of ASKθ, BES1, BZR1 and BEH2 were cloned into the binary plant expression vector, pGWR8, under the control of the 35S promoter. pGWR8, a derivative of pGreenII (Hellens et al., 2000 (link)), carries the translational leader of the tobacco etch virus for efficient translation. cDNAs were tagged with the Myc epitope or YFP. The constructs were transformed into Agrobacterium tumefaciensGV3101/pSoup by electroporation (Hellens et al., 2000 (link)). Transgenic A. thaliana Col-0 lines were generated by the floral-dip method (Clough and Bent, 1998 (link)), and were then selected on kanamycin-containing medium (50 μg ml⁻¹).

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Rozhon W., Mayerhofer J., Petutschnig E., Fujioka S, & Jonak C. (2010). ASKθ, a group-III Arabidopsis GSK3, functions in the brassinosteroid signalling pathway. The Plant Journal, 62(2), 215-223.

Publication 2010

Agrobacterium Bent Cdnas Electroporation Epitope Kanamycin Plant Tobacco etch virus Transgenic Vector

Corresponding Organization :

Other organizations : Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, RIKEN Advanced Science Institute

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Variable analysis

independent variables

CDNAs of ASKθ, BES1, BZR1 and BEH2

dependent variables

Transgenic A. thaliana Col-0 lines

control variables

35S promoter
Tobacco etch virus translational leader
Myc epitope or YFP tags
Agrobacterium tumefaciens GV3101/pSoup
Floral-dip method
Kanamycin-containing medium (50 μg ml^-1)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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