The complete sequence of theBacillus amyloliquefaciensproviral H2,BamHI methylase gene

We have previously isolated a gene from Bacillus amyloliquefaciens which encoded a DNA methylase with specificity for the BantHl site (1). Plasmid and genomic DNA isolated from E. coli HB101 cells harboring a plasmid pBamM2.5 containing this gene was resistant to cleavage by BamWl endonuclease. Expression of this methylase gene is dependent on orientation in pSP64 and under control of the LacZ promoter. The sequence analysis of this gene is shown below. Comparison of this sequence to a partial sequence of a Bacillus amyloliquefaciens prophage encoded methylase indicates that the gene we isolated encodes the prophage H2 BaniHl methylase (2). The sequence contains a 795 base pair open reading frame, nucleotides #441-1236 encoding a 265 amino acid, 30978.7 dalton protein. Deletion analysis of this clone suggests that this is the correct protein. A HindUl-HindlU DNA fragment, nucleotides #1 -651 , was subcloned into pSP64. Clones containing this deletion, which removes 196 carboxyl terminal amino acids, do not express BamHl methylase activity.