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:
Jun. 25, 2024

Filed:

Jul. 27, 2021
Applicant:

Wisconsin Alumni Research Foundation, Madison, WI (US);

Inventors:

Srivatsan Raman, Middleton, WI (US);

Phil Huss, Madison, WI (US);

Assignee:
Attorneys:
Primary Examiner:
Int. Cl.
CPC ...
C40B 50/06 (2006.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01);
U.S. Cl.
CPC ...
C12N 15/1093 (2013.01); C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/11 (2013.01); C12N 2310/20 (2017.05); C12N 2795/10052 (2013.01);
Abstract

Described herein is a method of preparing an unbiased library of phage variants, comprising (a) preparing a population of 'acceptor phage'; (b) removing an endogenous target gene and inserting gene variants into the acceptor phage genomes; (c) enriching the recombined phages; and (d) expressing the library for selection. The acceptor phage is a lytic phage comprising a synthetic genome wherein the target gene of interest is flanked by recombinase sites. The acceptor phage infects a first host bacteria expressing a recombination plasmid facilitating recombination. The phages then infect a second host bacteria expressing a counterselection system that accumulates recombined phage variants and selecting against non-recombined phages. The accumulated phage variants infect a third host bacteria. The phage library may then be sequenced and characterized.


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