Validation, Cloning, and Expression of Genes Involved in Histone Acetylation and Deacetylation in Common Bean (Phaseolus vulgaris L.)

Epigenetics is the study of heritable modifications in the functional activity of genes without changes in the DNA sequence. Our long-term goal is to better understand epigenetic control of important traits to help improve plant breeding and agriculture. Histone acetylation is a post-translational modification involved in gene regulation in eukaryotes. Acetyltransferases are enzymes that cause histone acetylation, which is associated with gene expression, while deacetylases cause deacetylation correlating to repression of genes. Common bean is used widely for its high protein and dietary fiber content.  To understand genes involved in acetylation and deacetylation of common bean, we obtained sequences from the GCN5 family of acetyltransferases and RPD3 family of deacetylases from the model plant, Arabidopsis thaliana. These sequences encode for genes involved in acetylation and deacetylation, respectively. The homologs of these genes were found from sequences of G19833, the reference common bean genotype in phytozome; a plant sequence database. Primers were designed to amplify DNA extracted from the leaves of Sierra; a rust resistant genotype of common bean, and G19833. Results suggest that the putative genes were amplified with both primers in Sierra. Full-length genes will be cloned for sequencing and the proteins will subsequently be expressed using an expression vector. The resulting proteins will be also used to produce antibodies for the preparation of chromatin immunoprecipitation to locate specific DNA regions bound by these proteins. This will lead to further understanding of GCN5 and RPD3 important families of plant proteins in common bean and potential use in epigenetics research.