Many crop plants, like soybean and canola, have pods that open to release seeds. However, the molecular mechanism of this pod-opening process is not well understood. I have studied two genes, called SCRAMBLED (SCM) and QUIRKY (QKY), that regulate this process in Arabidopsis thaliana. SCM encodes for a leucine-rich repeat receptor-like kinase and QKY encodes for a transmembrane protein with C2 domains. Mutations in these genes cause the plants to have small, twisted, and non-shattering pods contrasting to wild type pods, which are long, straight, and shattering. I carried out a double mutant analysis to determine if SCM and QKY genes function in the same genetic pathway. scm qky double mutants show very similar phenotypes to scm and qky phenotypes indicating that both genes function in the same genetic pathway. Using yeast two-hybrid assay, I showed that SCM and QKY proteins do not bind to each other in yeast. In addition, I screened a flower cDNA library using the yeast two-hybrid system for interactors of QKY, SCM extracellular domain, and SCM kinase domain. I found many interactors of SCM in yeast. However, so far, knock-out mutants of the genes did not show scm-like phenotypes. Further tests need to be done in order to determine if any of these putative interactors of SCM actually bind to SCM in Arabidopsis.Yeast transformation for testing protein interaction I used materials from Invitrogen ProQuest Two-Hybrid System with Gateway Technology (obtained from Dr. Jeff Long lab at Salk Institute) for all yeast transformations. Using the protocolanbsp;...
|Title||:||Finding Interactions of SCRAMBLED by Using the Yeast Two-hybrid System|
|Author||:||Jacqueline Phuong Anh Nguyen|
|Publisher||:||ProQuest - 2008|