Supplementary Figures
Arabidopsis tonoplast proteins TIP1 and TIP2 interact with the cucumber mosaic
virus 1a replication protein, by M. J. Kim, H. R. Kim and K.-H. Paek
Journal of General Virology vol. 87, part 11, pp. 3425-3431
Fig. S1. Interaction analyses of Arabidopsis tonoplast intrinsic proteins (TIPs)
and cucumber mosaic virus (CMV) or tobacco mosaiv virus (TMV) proteins using the yeast two-hybrid
system. (a) Interaction analyses of Arabidopsis TIPs and CMV 1a. cDNA encoding CMV 1a was
fused with the GAL4 activation domain in pGADT7, and different classes of cDNAs encoding
TIPs were fused with the GAL4 binding domain in pGBKT7. (b) Interaction analyses of
Arabidopsis TIPs and TMV replicase or helicase. cDNA encoding TMV replicase (nt 276-3419 of
TMV; GenBank accession no. V01408) or helicase (nt 2553-3323) was fused with the GAL4
activation domain. The two fusion vectors were co-transformed into the Y187 yeast strain, selected
on plates devoid of Leu and Trp, and then subjected to colony-lift filter assays using X-Gal as a
substrate. The pVA3-1/pTD1-1 and unfused pGADT7/pGBKT7 vectors were utilized as positive and
negative controls, respectively. The blue spots indicate β-galactosidase activity.
Fig. S2. Liquid assays for quantification of β-galactosidase activity using a
sensitive luminescent detection system (Clontech). The values are means±SD (n=3)
and are expressed as relative light units (RLU) normalized to cell density (OD600).
In the evaluation of each interaction, three separate transformed colonies were assayed and each of
the assays was performed in triplicate.
