Method for cloning and expression of SbfI restriction endonuclease and SbfI methylase in E. coli
View Patent ↗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.
1. Isolated DNA encoding the SbfI restriction endonuclease, wherein the isolated DNA is obtainable from Streptomyces species Bf-61.
2. A recombinant DNA vector comprising a vector into which a DNA segment encoding the SbfI restriction endonuclease has been inserted.
3. Isolated DNA encoding the SbfI restriction endonuclease and SbfI methylase, wherein the isolated DNA is obtainable from ATCC No. PTA-5371.
4. A vector which comprises the isolated: DNA of claim 3 .
5. A host cell transformed by the vector of claim 2 or 4 .
6. A method of producing recombinant SbfI restriction endonuclease comprising culturing a host cell transformed with the vector of claim 2 or 4 under conditions suitable for expression of said endonuclease.