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: RsaI, a restriction enzyme from the bacterium Rhodopseudomonas sphaeroides, recognizes the DNA sequence 5′-GTAC-3′. Because RsaI is commercially valuable, we sought to overproduce it by cloning the genes for RsaI and its accompanying, modification, enzyme. The ‘methylase-selection’ method, the customary procedure for cloning restriction and modification genes, was applied to RsaI. The method yielded clones containing the methylase gene (rsaIM), but none containing both the methylase gene and the restriction gene (rsaIR). Inverse-PCR was then used to recover sections of the DNA downstream of rsaIM. These sections were sequenced, and the sequences were joined in silico to reveal the gene organization of the RsaI R-M system.

Abstract: The methylase selection method was used to clone the SnaBI methylase gene (snaBIM) from Sphaerotilus natans (ATCC 15291). An active SnaBI methylase was cloned in E. coli using pSnaBI-2, a pUC19 derivative containing two SnaBI sites. Because methylase and restriction genes are usually located alongside each other in a restriction-modification systems, efforts were made to clone the downstream DNA by inverse PCR. Inverse PCR cloned the missing portion of the SnaBI endonuclease and also identified a control, or C, protein. The two open reading frames snaBIR (ORF1) and snaBIC (ORF2) converged toward the SnaBI methylase gene (ORF). Attempts to establish a snaBIR-recombinant plasmid expressing the SnaBI endonuclease in E.coli modified with SnaBI methylase failed. Overexpression of the SnaBI endonuclease in E. coli required the use of the heterospecific BsaAI methylase.

Abstract: A recombinant restriction endonuclease from Anabaena variabilis is provided, as well as the isolated gene which encodes it and methods for the production of the recombinant AvaI restriction endonuclease. An isolated gene encoding a modification methylase from A. variabilis is also provided.

Abstract: The present invention relates to recombinant DNA which encodes the AgeI restriction endonuclease as well as AgeI methyltransferase, and production of AgeI restriction endonuclease from E. coli cells containing the recombinant DNA.

Abstract: In accordance with the present invention there is provided an isolated DNA coding for the Bg/I restriction endonuclease and modification methylase derived from Bacillus globigii RUB561 stain, as well as related methods for cloning said recombinant DNA. The present invention also relates to clones which express recombinant Bg/I restriction endonuclease and recombinant modification methylase produced from Bg/I recombinant DNA and to methods for producing said enzymes.

Abstract: The present invention is directed to a method for cloning and producing the MwoI restriction endonuclease by 1) introducing the restriction endonuclease gene from M. wolfei into a host whereby the restriction gene is expressed; 2) fermenting the host which contains the plasmid encoding and expressing the MwoI restriction endonuclease activity, and 3) purifying the MwoI restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the MwoI restriction endonuclease activity.

Abstract: The present invention is directed to a method for cloning and producing the Afl II restriction endonuclease by 1) introducing the restriction endonuclease gene from Anabaena flos-aquae into a host whereby the restriction gene is expressed; 2) fermenting the host which contains the plasmid encoding and expressing the Afl II restriction endonuclease activity, and 3) purifying the Afl II restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the Afl II restriction endonuclease activity.

Abstract: The present invention is directed to a method for cloning and producing the XmaI restriction endonuclease by (1) introducing the restriction endonuclease gene from X. malvacaerum into a host whereby the restriction gene is expressed; (2) fermenting the host which contains the plasmid encoding and expressing the XmaI restriction endonuclease activity, and (3) purifying the XmaI restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the XmaI restriction endonuclease activity.

Abstract: The present invention is directed to a method for cloning and producing the HgiAI restriction endonuclease by (1) introducing the restriction endonuclease gene from Herpetosiphon giganteus into a host whereby the restriction gene is expressed; (2) fermenting the host which contains the vector encoding and expressing the HgiAI restriction endonuclease, and (3) purifying the HgiAI restriction endonuclease from the fermented host which contains the vector encoding and expressing the HgiAI restriction endonuclease activity.