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. 2008 Apr 25;54(6):1083–1093. doi: 10.1111/j.1365-313X.2008.03481.x

Figure 2.

Figure 2

Interaction assays of LeCTR2 with LeETR1 and NR in the yeast two-hybrid system. (a) Structures of LeETR1, NR and LeCTR2, with residues numbered. (b) Constructs used for protein–protein interaction assays in yeast. (c) Yeast expressing bait proteins in the absence of the prey constructs was tested for activation of the LacZ reporter gene by incubation on minimal medium containing glucose (Glu) and X-gal (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside). (d) Synthesis of bait proteins in yeast was detected by Western blotting using an anti-LexA antibody. Lane 1, DB–ETR1132–754; lane 2, DB–ETR1HKR364–754; lane 3, DB–ETR1HK364–647; lane 4, DB–NR117–635; lane 5, DB–ETR1R647–754. Mw, molecular weight markers. (e) Yeast expressing DB–ETR1132–754/AD–CTR2N50–700, DB–NR117–635/AD–CTR2N50–700, DB–ETR1132–754/AD–CTR2C680–982, DB–NR117–635/AD–CTR2C680–982, DB–ETR1HKR364–754/AD–CTR2N50–700, DB–ETR1HK364–647/AD–CTR2N50–700 or DB–ETR1R647–754/AD–CTR2N50–700 was tested for LacZ reporter gene expression on minimal medium containing X-gal in the presence of galactose (Gal/X-gal) or glucose (Glu/X-gal).