Protein phosphatase 4 recombinant protein – transformation of bacteria and Arabidopsis thaliana

Abstract

Protein phosphatase 4 (PP4) is well conserved in eukaryote organisms. The protein has been studied in yeast and mammalian cells, but little is known about PP4 in plants. The function of PP4 in plants is unknown and it will not have the same function as in the other studied organisms because the processes does not exist in plants. It is likely that the PP4 proteins has been adapted for other uses in plants. In this thesis project, I worked toward making Arabidopsis thaliana plants with the two PP4 proteins PP4-1 and PP4-2 using a variety of recombination technologies. Plants were successfully transformed with PP4-2 and PP4-1. The plants with PP4-2 made seeds that have a chance of having the PP4-2 gene themselves. These plants can be used for further study of PP4-2 in plants. As the practical part of the thesis work ended, the plants transformed with the PP4-1 gene were starting to produce seeds that later can be selected for the gene and grow to produce more seeds. These can potentially be used in further studies. I also demonstrated that the PP4-2 protein could be produced with E. coli Rosetta strain. The protein was visualized on a polyacrylamide gel. The PP4-1 has been successfully cloned into a pMAL-C2X vector that can be used for further studies.