Strand-specific polynucleotide nickases

A strand-specific polynucleotide nickase comprising a heteromeric restriction endonuclease which comprises a first subunit and a second subunit and which recognises an asymmetric nucleotide recognition sequence, wherein the first subunit comprises a catalytic domain capable of cleaving one strand of a DNA duplex, and the second subunit is modified to render the catalytic domain thereof inactive.

A strand-specific polynucleotide nickase comprising a R.Bpu 10I heteromeric restriction endonuclease which comprises a first subunit and a second subunit and which recognises an asymmetric nucleotide recognition sequence, wherein the first subunit comprises a catalytic domain capable of cleaving one strand of a DNA duplex, and the second subunit is modified to render the catalytic domain thereof inactive.

A process for producing a strand-specific polynucleotide nickase, which process comprises inactivating the catalytic activity of one subunit of a restriction endonuclease, wherein the endonuclease comprises a first subunit comprising a catalytic domain capable of cleaving one strand of a DNA duplex and a second subunit comprising a catalytic domain capable of cleaving the other strand of the DNA duplex, and the endonuclease recognises an asymmetric nucleotide recognition sequence.

A process for producing a strand-specific polynucleotide nickase, which process comprises inactivating the catalytic activity of one subunit of a heteromeric restriction endonuclease, wherein the endonuclease comprises a first subunit comprising a catalytic domain capable of cleaving one strand of a DNA duplex and a second subunit comprising a catalytic domain capable of cleaving the other strand of the DNA duplex, and the endonuclease recognises an asymmetric nucleotide recognition sequence.

A process for producing a strand-specific polynucleotide nickase, which process comprises inactivating the catalytic activity of one subunit of a heteromeric restriction endonuclease R.Bpu10I.