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. 2019 Mar 4;20(5):1104. doi: 10.3390/ijms20051104

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Identification of prey proteins interacting with TaGAPCp1. (a) Confirmation of true positive clones by small-scale Y2H assay. First row and second row: pGBKT7-TaGAPCp1 plasmid and the respective prey 1 to prey 7 plasmids (P1–P7) was co-transformed into Y2HGold Yeast Strain then staked on QDO/X/A plates and DDO/X/A plates, respectively. Third row and fourth row: pGBKT7-TaGAPCp1 plasmid and pGBKT7 empty bait was co-transformed into Y2HGold Yeast Strain then staked on QDO/X/A plates and DDO/X/A plates, respectively. PC indicates a positive control, NC indicates a negative control. (b) BiFC assay of the interaction between TaGAPCp1 and Cyt b6f proteins in tobacco leaf protoplasts. The pSPYNE-TaGAPCp1 and pSPYCE-Cyt b6f constructs were co-infiltrated in tobacco by Agrobacterium. The YFP fluorescence was detected by confocal laser scanning microscopy. Co-transformants of pSPYNE-TaGAPCp1 and pSPYCE as well as pSPYNE and pSPYCE-Cyt b6f were used as negative controls. Bar = 20 μm.

Identification of prey proteins interacting with TaGAPCp1. (a) Confirmation of true positive clones by small-scale Y2H assay. First row and second row: pGBKT7-TaGAPCp1 plasmid and the respective prey 1 to prey 7 plasmids (P1–P7) was co-transformed into Y2HGold Yeast Strain then staked on QDO/X/A plates and DDO/X/A plates, respectively. Third row and fourth row: pGBKT7-TaGAPCp1 plasmid and pGBKT7 empty bait was co-transformed into Y2HGold Yeast Strain then staked on QDO/X/A plates and DDO/X/A plates, respectively. PC indicates a positive control, NC indicates a negative control. (b) BiFC assay of the interaction between TaGAPCp1 and Cyt b6f proteins in tobacco leaf protoplasts. The pSPYNE-TaGAPCp1 and pSPYCE-Cyt b6f constructs were co-infiltrated in tobacco by Agrobacterium. The YFP fluorescence was detected by confocal laser scanning microscopy. Co-transformants of pSPYNE-TaGAPCp1 and pSPYCE as well as pSPYNE and pSPYCE-Cyt b6f were used as negative controls. Bar = 20 μm.