Heterologous expression and purification of Arabidopsis thaliana VIM1 protein: In vitro evidence for its inability to recognize hydroxymethylcytosine, a rare base in Arabidopsis DNA

► Expression and purification of the full-length Arabidopsis VIM1 protein in Escherichia coli. ► First report of 5-hydroxylmethylcytosine (5hmC) in Arabidopsis genomic DNA. ► Characterize binding of recombinant VIM1 to synthetic DNAs with 5mC and 5hmC. ► Results show that 5hmC could drastically reduce VIM1-mediated methylation of replicated DNA.