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. 2020 Feb 6;25(3):700. doi: 10.3390/molecules25030700

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© 2020 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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Yeast-two hybrid analysis of TCMP-1 interaction with heavy metal-associated isoprenylated plant protein 26-like (SlHIPP26). Yeast cells transformed with different combinations of constructs containing TCMP-1 fused with the DNA binding domain (BD) (bait; TCMP-1), HIPP26 fused with the activation domain (AD) (prey; HIPP26), the BD alone (bait; empty), and the AD alone (prey; empty) were grown on two different selective media. Interaction of p53 with SV40 was used as a positive control of the mating system. (a) The mated cultures were spotted on SD/-Leu/-Trp control plates to confirm the presence of both plasmids in yeast cultures. (b) The mated cultures were spotted on synthetic quadruple dropout agar plates (SD/-Leu/-Trp/-His/-Ade/X-Gal/Aureobasidin A) to check genuine positive interactions. Positive interaction was confirmed by the growth of blue colonies (◣, indicates that two increasing dilutions of the mated cultures were used).