Validation of protein-protein interaction among a selected set of putative MADS box interactors by yeast-two-hybrid technique
Student name: Ms Ridhi Khurana
Guide: Dr Ramakrishnan Sitaraman
Year of completion: 2013
Host Organisation: Department of Plant Molecular Biology, University of Delhi South Campus
Supervisor (Host Organisation): Dr Sanjay Kapoor
Abstract: MADS box is a conserved DNA-binding domain found conserved among four genes across four
species, namely, Mcm1 of Saccharomyces cerevisiae, AGAMOUS of Arabidopsis, DEFICIENS of
Antirrhinum and SRF (Serum Response Factor) of humans. Two separate groups identified it for
the first time in plants in 1990, Theiβen et al. in Arabidopsis thaliana and Schwarz-Sommer et al.
in Antirrhinum majus. MADS box genes are similar to the homeobox or hox genes of animals as
they are both found to be responsible for organ development. MADS box gene family is well
known for the role of its members in floral transition and development. Also, over the recent
years their role has been elucidated in development of plant organs apart from those in the
flower such as, root development, tillering, pollen maturation, fruit ripening, endosperm
development, endothelium development and seed development. The entire MADS box gene family
in rice, comprising of 75 genes was identified based on computational and bioinformatic
methods by Arora et al. (2007). The present study revolves around the role of MADS box genes
in seed development in rice. It is aimed at examining their interactions using the yeast twohybrid
technique.A number of MADS box genes cloned into pENTR/D-TOPO vectors based on
previous knowledge from microarrays and real time expression analysis data were provided.
These were mobilized into yeast two-hybrid destination vectors via GATEWAY cloning. Bait and
Prey constructs were generated using this method and transformed into yeast strains with
opposite mating types for one to one mating studies and co-transformed into a single strain for
co-transformation based interaction studies. The results for the co-transformation yielded
positive interaction between the bait and the prey construct for one of the clones under study,
indicating towards its homodimerization.
Keywords: Rice, MADS Box, Seed Development, Yeast Two –Hybrid, ProteinProtein
Interactions.
