Cloned KpnI restriction-modification system

Details Cloned KpnI restriction-modification system Cloned KpnI restriction-modification system

1990-03-20

1992-01-21

Patterson, Charles L.

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

4353201, 435199, 435193, 536 27, C12N 121, C12N 1570, C12N 1555, C07H 1910

Patent

active

050827847

ABSTRACT:
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.

REFERENCES:
Tomassini, T. et al., (1978), Nuc. Acids Res. 5(11), 4055-4064.
Mann et al., Cloning of Restriction and Modification Genes in E. coli: The HhaII System from Haemophilus Haemolyticus, Gene, 3:97-112 (1978).
Brooks et al., Cloning the BamHI Restriction Modification System, Nucleic Acids Research, 17:(3) 979-997 (1989).
Howard et al., Cloning the DdeI Restriction-Modification System Using a Two-Step Method, Nucleic Acids Research, 14:(20) 7939-7951 (1986).
Szomolanyi et al., Cloning the Modification Methylase Gene of Bacillus Sphaericus R in Escherichia coli, Gene, 10:219-225 (1980).
Wilson, G., Cloned Restriction-Modification Systems-A Review, Gene, 74:281-289 (1988).
Rodicio et al., Cloning and Expression of the Sa1I Restriction-Modification Genes of Streptomyces Albus G, Mol. Gen. Genet., 213: 346-353 (1988).

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