Yeast-2-Hybrid Protein Analysis of Arabidopsis Thaliana Trichomes

Abstract

Arabidopsis thaliana trichomes are specialized single-cellular structures on the surface of leaves. They have a unique, three dimensional, branching structure and can become morphologically mutated as the results of many genetic changes. One gene known to function in trichome development is At1g64690, also known as AtBLT. AtBLT contains a region of DNA that could form a coiled coil with a region that should also form a coiled coil in another gene. To identify genes that may interact with this region of AtBLT that also act in trichome development, yeast two hybrid analysis will be used. BLT will be expressed in a vector containing a DNA binding domain; and the gene of interest, in this case STIIA, will be expressed in a vector containing a DNA activation domain. The two constructs, acting as bait and prey, are expressed in yeast with a Gal reporter that will cause the yeast to metabolize beta-galactosidase and turn a blue color if the two proteins interact by forming a coiled coil and bringing the binding domain and activation domain together.