Abstract: An isolated DNA is provided which encodes a protein that is capable of binding to 5?GGTACC-3?, the isolated DNA being capable of hybridizing to SEQ ID NO:3 under stringent hybridization conditions. The isolated DNA may be alternatively characterized as coding for a protein having an amino acid sequence comprising SEQ ID NO:5 or by an amino acid sequence with an expectation value of less than E=e?02 in a BLAST search using SEQ ID NO:5. Vectors containing the isolated DNA and host cells expressing the vectors as well as a method for making recombinant Acc65I having the above properties are also provided.

Abstract: An isolated DNA is provided which encodes a protein that is capable of binding to 5?GGTACC-3?, the isolated DNA being capable of hybridizing to SEQ ID NO:3 under stringent hybridization conditions. The isolated DNA may be alternatively characterized as coding for a protein having an amino acid sequence comprising SEQ ID NO:5 or by an amino acid sequence with an expectation value of less than E=e?02 in a BLAST search using SEQ ID NO:5. Vectors containing the isolated DNA and host cells expressing the vectors as well as a method for making recombinant Acc65I having the above properties are also provided.

Abstract: A novel restriction endonuclease and methods of making the same are obtainable from either Citrobacter species 2144 (NEB#1398) or the recombinant stain Escherichia coli (NEB#1554) which cleaves at nt sequence 5?-CAANNNNNGTGG-3? (SEQ ID NO:14) in double-stranded DNA molecules. The novel restriction endonuclease is a modular protein in which the specificity moiety is an independent module from the restriction-modification module.

Abstract: Methods are provided for converting into a sequence specific strand specific and location specific DNA nicking endonuclease, a restriction endonuclease that recognizes an asymmetric DNA sequence, the endonuclease having two catalytic sites and one or more single sequence specific DNA-binding domains. In one embodiment the method requires inactivating one of the catalytic sites of the restriction endonuclease. In another embodiment, the restriction endonuclease is a dimer having a first and second subunit each comprising a sequence specific DNA binding domain, a catalytic site and a dimerization domain. The nicking endonuclease is formed from combining one subunit having an inactivated catalytic site and a second subunit having an inactivated DNA binding domain. The nicking endonuclease may be converted into a restriction endonuclease by the addition of manganese cations in the digestion buffer.

Abstract: The present invention relates to: recombinant DNA encoding the SbfI restriction endonuclease as well as the SbfI methylase, and expression of the SbfI restriction endonuclease and SbfI methylase in E. coli cells containing the recombinant DNA; and methods for cloning the SbfI restriction gene (sbfIR) from Streptomyces species Bf-61 into E. coli by PCR. The method relied on primers based on DNA sequences predicted from amino acid sequences of the purified SbfI restriction endonuclease.

Abstract: Methods are provided for converting into a sequence specific strand specific and location specific DNA nicking endonuclease, a restriction endonuclease that recognizes an asymmetric DNA sequence, the endonuclease having two catalytic sites and one or more single sequence specific DNA-binding domains. In one embodiment the method requires inactivating one of the catalytic sites of the restriction endonuclease. In another embodiment, the restriction endonuclease is a dimer having a first and second subunit each comprising a sequence specific DNA binding domain, a catalytic site and a dimerization domain. The nicking endonuclease is formed from combining one subunit having an inactivated catalytic site and a second subunit having an inactivated DNA binding domain. The nicking endonuclease may be converted into a restriction endonuclease by the addition of manganese cations in the digestion buffer.

Abstract: The present invention provides a novel type II restriction endonuclease obtainable from Arthrobacter protophormiae. The endonuclease known as Apo I, recognizes the following nucleotide sequence and has a cleavage point indicated by the arrows: ##STR1## Also described is a process for obtaining Apo I from Arthrobacter protophormiae.