Cloning the SOS4 gene

Abstract

The SOS4 (Salt overly sensitive 4) gene encodes a pyridoxal kinase that is involved in the biosynthesis of pyridoxal-5-phosphate, an active form of vitamin B6. Vitamin B6 plays an important role in cell metabolism. We utilized Polymerase Chain Reaction (PCR) to amplify the SOS4 from cDNA. This product was then ligated into pGBKT7, a vector used in a Yeast 2-Hybrid protein interaction system, and transformed into Escherichia coli. After screening this construct for mutations, we will transform the construct into Saccharomyces cerevisiae and perform a Yeast 2-Hybrid screen, in which we will look for proteins that interact with SOS4.