We used the ProQuest Two-Hybrid System (Invitrogen) as previously described (28 (link)). Specifically, the cDNA encoding full-length GI was transferred from the pENTR/D-TOPO vector (Invitrogen) into the pDEST32 vector by Gateway LR recombination reaction (Invitrogen) to generate the bait plasmid; the pDEST22 prey plasmids containing the sequences encoding RGA, GAI, RGL1, RGL2, and RGL3 have been previously described (45 (link)). Empty pDEST22 and the pExpAD502 plasmids were used as negative controls. Quantitation of β-galactosidase activity was performed in a 96-well format as previously described (47 (link)).
Free full text: Click here