pSAT-GalBD:vp16 plasmid vector質粒載體

Transcriptional activation assay in yeast cell. Yeast co-transformants were incubated on the selective medium (SD/-Ade/-His/-Leu/-Trp plus X-α-Gal) at 30?°C for 3 d. If TH1 is a transcription activator, the β-Galactosidase reporter gene will be expressed and the filter will be stained blue in the Colony-lift Filter Assay. pGADT7-RecT & pGBKT7-53 (positive control); pGADT7-RecT & pGBKT7-Lam (negative control). (b) Constructs used in transient expression assay in rice leaf protoplasts. Six different effector plasmids were made by fusing TH1, TH1S2-89 with activator domain VP16, repressor domain EAR or the GAL4 DNA binding domain (GAL4-DB). The reporter plasmid Pro35S-GAL4:LUC contains a CaMV 35?S promoter, 5?×?GAL4 binding site, a TATA box in front of the firefly luciferase (LUC) gene and a nopaline synthase terminator. The p35S: REN plasmid expressing the Renilla luciferase was used as an internal control. (c) Relative luciferase activities in rice leaf protoplasts using TH1, TH1s2-89, VP16, TH1-VP16, TH1s2-89-VP16, EAR, TH1-EAR and TH1s2-89-EAR as effector compared with GAL4-DB control. Error bars indicate SE (n?=?3). Double asterisks indicate significant difference at P?<?0.01 by Student’s t test.

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