human fetal brain matchmaker cdna library  (TaKaRa)

Journal: PLoS ONE

Article Title: Direct Interaction of Selenoprotein R with Clusterin and Its Possible Role in Alzheimer’s Disease

doi: 10.1371/journal.pone.0066384

Figure Lengend Snippet: Plasmids carrying on the fetal brain cDNA library were co-transformed into the NpGBKT7- SelR′ -containing yeast and screened by the selection plate for the blue colonies (A). The interaction between SelR′ and Clu was verified by re-transformation of the plasmids NpGBKT7- SelR′ and pACT2- Clu into either AH109 (B) or Y2HGold (C) yeast cells. Yeast cells in B (1–3) were transformed with single NpGBKT7- SelR′ , NpGBKT7- SelR′ plus pACT2, and NpGBKT7- SelR′ plus pACT2- Clu plasmids, respectively. Yeast cells in C, D, E were transformed with NpGBKT7- SelR′ plus pACT2- Clu , pGBKT7- p53 plus pADT7- T (positive control), and pGBKT7- Lam plus pADT7- T (negative control), respectively, followed by the selection on SD/−Leu/−Trp/X-α-Gal/Aba plates.

Article Snippet: Matchmaker™ Gold yeast two-hybrid system, yeast strains Y2HGold and AH109, plasmids pACT2 and NpGBKT7, and human fetal brain cDNA library (using pACT2 as the vector) were purchased from Clontech Laboratories (Mountainview, CA, USA).

Techniques: cDNA Library Assay, Transformation Assay, Selection, Positive Control, Negative Control

Journal: PLoS ONE

Article Title: GRG5/AES Interacts with T-Cell Factor 4 (TCF4) and Downregulates Wnt Signaling in Human Cells and Zebrafish Embryos

doi: 10.1371/journal.pone.0067694

Figure Lengend Snippet: The AH109 yeast strain was co-transformed with the indicated constructs (central panel). Transformants were selected in medium lacking tryptophan and leucine (left panel) and activation of a Gal4-dependent ADE2 gene was examined by growth on selective plates additionally lacking adenine (right panel). Yeast cells containing both TCF4 and GRG5/AES grew in the selective media. pLAM and pACT2 plasmids were used as bait and prey negative controls respectively, while the β-catenin prey plasmid constitutes a positive control for the interaction assays.

Article Snippet: A commercial human fetal brain cDNA library in pACT2 (Clontech), was screened by PEG/LiAcetate co-transformation [ ] with the bait plasmid pAS2-TCF4 into the AH109 yeast reporter strain (which contains integrated copies of ADE2 and HIS3 reporter genes under control of Gal4-dependent promoters).

Techniques: Transformation Assay, Construct, Activation Assay, Plasmid Preparation, Positive Control