Cloning and Nucleotide Sequence ofApaLI Restriction-modification System fromAcetobacter pasteurianusIFO 13753†

The ApaLI restriction-modification system from Acetobacter pasteurianus IFO 13753 recognizes the nucleotide sequence GTGCAC. The gene coding for the ApaLI methylase (M.ApaLI) was cloned into Escherichia coli DH5 alpha MCR, and the nucleotide sequence of the gene was analyzed. The M.ApaLI gene coded for a protein of 429 amino acid residues (molecular mass, 46,554 daltons). The ApaLI restriction endonuclease (R.ApaLI) gene was analyzed by inverse polymerase chain reaction. The R.ApaLI gene coded for a protein of 375 amino acid residues (molecular mass, 42,143 daltons). The two genes had the same orientation separated by two base pairs. The deduced amino acid sequence of M.ApaLI shows significant similarities to the family of cytosine-5 methylases. However, the deduced amino acid sequence of R.ApaLI did not have as much relatedness in the nucleotide sequence, when compared with those of the other restriction endonucleases already reported.