Cloned kpni restriction-modification system

Details Cloned kpni restriction-modification system Cloned kpni restriction-modification system

C - Chemistry, Metallurgy

12

N

C12N 15/55 (2006.01) C12N 1/21 (2006.01) C12N 9/10 (2006.01) C12N 9/16 (2006.01) C12N 9/22 (2006.01) C12N 15/54 (2006.01) C12N 15/70 (2006.01)

Patent

CA 2078554

2078554 9114771 PCTABS00007 The present invention discloses the cloning and expression in a host such as Escherichia coli of the KpnI restriction-modification system from Klebsiella pneumoniae, utilizing a two step protocol. Initial protection of the E. coli host with methylase expressed on a vector was required to stabilize a compatible vector carrying both the endonuclease and the methylase genes on a single DNA fragment. A chromosomal map was generated localizing the genes for KpnI methylase and endonuclease. An E. coli) strain was constructed which produced several thousand-fold higher levels of KpnI endonuclease than the level produced by Klebsiella pneumoniae. This invention is also directed to cloning and expression of genes encoding for restriction endonuclease isoschizomers of KpnI and/or modification methylase isoschizomers of KpnI methylase.

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