The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
May. 17, 2011

Filed:

Dec. 15, 2004
Applicants:

Shuang-yong Xu, Lexington, MA (US);

James Samuelson, Newburyport, MA (US);

Inventors:

Shuang-yong Xu, Lexington, MA (US);

James Samuelson, Newburyport, MA (US);

Assignee:

New England Biolabs, Inc., Ipswich, MA (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12Q 1/00 (2006.01); C12N 15/00 (2006.01); C12N 15/09 (2006.01); C12N 15/11 (2006.01); C12N 15/52 (2006.01);
U.S. Cl.
CPC ...
Abstract

Methods are provided for engineering novel strand-specific nicking endonucleases by means of an in vivo enrichment of a plasmid library containing a randomly mutagenized restriction endonuclease gene. The plasmids contain adjacent to the gene a cleavable or nickable sequence for cleaving or nicking by the endonuclease product of the gene and a second recognition site for a second endonuclease. The plasmid library is used to transform unmodified host cells. Plasmids from the cultured transformed cells may be analyzed by an in vitro assay for nicking and the nicked plasmids pooled and used to transform host cells. The product is then pooled and the single-stranded specificity of the endonuclease is then determined. The product is either cloned after amplification or identified by use of a selectable marker.


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