Host genes involved in Agrobacterium-mediated transformation

Abstract

Agrobacterium is the natures genetic engineer that can transfer genes across the kingdom barriers to both prokaryotic and eukaryotic host cells. The host genes which are involved in Agrobacterium-mediated transformatiom (AMT) are not well known. Here, I studied in a systematic way to identify the whole eukaryotic host genes which are involved in this event. For this, I used a collection of around 4800 homozygous diploid deletion mutants and the cDNA libraries of yeast Saccharomyces cerevisiae as the eukaryotic host and two strains of Agrobacterium. First, a large-scale AMT protocol for the transformation of yeast mutants was developed. Then, the yeast mutant collection was screened for the mutants with an increased or a decreased AMT. 249 deletion mutant strains were found which affect AMT. Of great importance were host chromatin remodeling and histone modifying complexes. Besides, using GFP technique I showed the nuclear localization of Agrobacteriums VirD2 in yeast. Yeast two-hybrid screens also identified 12 yeast putative interaction partners of VirD2 protein.