Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1992-01-17
1993-03-09
Patterson, Jr., Charles L.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435193, C12N 922
Patent
active
051926750
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:
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Chatterjee Deb K.
Hammond Alan W.
Life Technologies Inc.
Patterson Jr. Charles L.
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