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. Author manuscript; available in PMC: 2008 Dec 13.
Published in final edited form as: J Mol Biol. 2007 Dec 23;376(4):1091–1099. doi: 10.1016/j.jmb.2007.12.032

Fig. 2.

Fig. 2

Y2H interaction analysis of wild-type CD-Dnmt3a and its variants with Dnmt3L. In the Y2H system, the CD-Dnmt3a/Dnmt3L interaction induces expression of HIS3 and lacZ reporter genes. HIS3 expression was tested by growing the yeast clones in a selective medium lacking histidine, and lacZ expression was analyzed by the β-galactosidase filter assay that is shown here. (a) Interaction of wild-type CD-Dnmt3a and Dnmt3L. All yeast colonies were positive in the β-galactosidase filter assay (indicating interaction of CD-Dnmt3a and Dnmt3L). Control experiments with the pACT2 vector and Dnmt3L did not show a β-galactosidase signal. Only a few clones were positive, that is, if the interaction of the CD-Dnmt3a random mutant library and Dnmt3L was tested. (b) Two examples of β-galactosidase filter assays performed with the interacting clones obtained after randomization and selection. (c) The CD-Dnmt3a F768E mutant disrupted the interaction with Dnmt3L as shown by the absence of β-galactosidase signal indicated by the absence of blue coloring.