Table 1 Vector construction used for the yeast two-hybrid screening and other assays by [5]
From: Potential important roles and signaling mechanisms of YPEL4 in pulmonary diseases
Type of assay | Final vector | Vector construction |
|---|---|---|
Yeast two-hybrid screening | pGBKT7-YPEL4 | Full-length cDNA of YPEL4 was ligated in frame with GAL4 DNA-binding domain of the pGBKT7 vector |
Mammalian two-hybrid reporter assay | YPEL4 expression vector: pCMV-BD-YPEL4 | Full-length cDNA of YPEL4 was cloned into mammalian expression plasmid pCMV-BD |
MVP expression vector: pCMV-AD-MVP | cDNA fragment of MVP was cloned into mammalian expression plasmid pCMV-AD | |
Co-localization assay | Vector EGFPN1 was used to construct vectors expressing YPEL4, and vector pCMV-Tag2B was used to construct a vector expressing MVP | |
GST pull-down assay | pGEX-4T-1 was used to construct vectors expressing GST-YPEL4 fusion proteins | |
Immunoprecipitation assay | pCMV-Tag2B and pCMV-Myc were used to construct vectors expressing YPEL4 and MVP, respectively | |