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Fig. 2 | BMC Plant Biology

Fig. 2

From: The walnut transcription factor JrGRAS2 contributes to high temperature stress tolerance involving in Dof transcriptional regulation and HSP protein expression

Fig. 2

Identification of the up-steam regulator of JrGRAS2 using yeast one-hybrid assays. a, Diagram of the reporter and effecter vectors. Three tandem copies of the DOFCOREZM motif were inserted into the pHis2 vector and used as the reporter construct. The CDS of JrDof3 was cloned into pGADT7-Rec2 and used as the effecter construct. b, The effecter and reporter constructs were co-transformed into the yeast strain Y187. p53His2 + pGADT7-p53, positive control, pGADT7-Rec2 vector that encodes murine p53 fused with GAL4 AD; p53His2 + pGADT7-JrDof3, negative control, pHis2 reporter vector that contains the cis-acting DNA consensus sequence recognized by p53. pHis2-DOF + pGADT7-JrDof3, pHis2-DOF-M + pGADT7-JrDof3, pHis2-DOF-S + pGADT7-JrDof3, pHis2-DOF-M1 + pGADT7-JrDof3, pHis2-DOF-M2 + pGADT7-JrDof3 means the interaction DOFCOREZM motif, mutated DOFCOREZM motif, promoter segment including the DOFCOREZM motif, promoter segment containing the mutated DOFCOREZM motif, promoter segment excluding the DOFCOREZM motif with JrDof3, accordingly. The transformants spotted on TDO plates with 0 mM 3-AT were used as positive controls for transformants growth. Positive transformants were further confirmed by spotting serial dilutions (1/1, 1/10, 1/100, 1/1000, 1/10000) onto TDO plates with 50 mM 3-AT

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BMC Plant Biology

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