Journal: Microbiology Spectrum

Article Title: Identification of a Novel Interaction between Theileria Prohibitin ( Ta PHB-1) and Bovine RuvB-Like AAA ATPase 1

doi: 10.1128/spectrum.02502-22

Figure Lengend Snippet: Yeast two-hybrid library screening. (A) Western blot showing the expression of myc-tagged Ta PHB-1 in Y2HGold yeast cells. (B) The presence of white colonies on SD/-Trp/X-α-Gal agar plate and no colonies on SD/-Trp/X-α-Gal/AbA plates confirmed the absence of the auto-activation of reporter genes by Ta PHB-1 in Y2HGold yeast cells. (C) Plates showing Y2HGold cells that grew on QDO/X/A agar plates after library screening. (D) Agarose gel electrophoresis analysis of PCR products of prey plasmids isolated from the colonies that grew on QDO/X/A agar plates. M, marker. (E) Growth of Y2HGold yeast cells on QDO/X/A agar plates after cotransformation of potential prey plasmid and pGBKT7- Ta PHB-1 confirmed their interaction. Cotransformation with pGBKT7-53 and pGADT7-T plasmids was used as a positive control, and cotransformation with pGBKT7-LAM and pGADT7-T plasmids was used as a negative control on the DDO/X/agar plate.

Article Snippet: The recombinant pGBKT7- Ta PHB-1 was transformed into a Y2HGold yeast strain following the Yeastmaker yeast transformation system 2 (TaKaRa).

Techniques: Library Screening, Western Blot, Expressing, Activation Assay, Agarose Gel Electrophoresis, Isolation, Marker, Plasmid Preparation, Positive Control, Negative Control