| US 7,563,578 B2 | ||
| Method for in vitro molecular evolution of protein function | ||
| Roland Carlsson, Lund (Sweden); Ann-Christin Malmborg Hager, Helsingborg (Sweden); Christina Furebring, Lund (Sweden); and Carl Borrebaeck, Hjarup (Sweden) | ||
| Assigned to Alligator Bioscience AB, Lund (Sweden) | ||
| Filed on Sep. 20, 2007, as Appl. No. 11/858,429. | ||
| Application 11/858429 is a continuation of application No. 11/185044, filed on Jul. 20, 2005, granted, now 7,282,334. | ||
| Application 11/185044 is a continuation of application No. 09/734801, filed on Dec. 12, 2000, granted, now 6,958,213. | ||
| Prior Publication US 2008/0248533 A1, Oct. 09, 2008 | ||
| This patent is subject to a terminal disclaimer. | ||
| Int. Cl. C12Q 1/68 (2006.01); C12P 19/34 (2006.01); C07H 21/04 (2006.01) | ||
| U.S. Cl. 435—6 [435/91.2; 536/24.33] | 6 Claims |
| 1. A method for generating a polynucleotide sequence or population of sequences from parent single stranded polynucleotide
sequences encoding one or more protein motifs, comprising the steps of
a) providing single stranded DNA constituting plus and minus strands of parent polynucleotide sequences;
b) digesting the single stranded polynucleotide sequences with a nuclease other than DNase I to generate populations of single
stranded fragments;
c) contacting said fragments generated from the plus strands with fragments generated from the minus strands and optionally,
adding primer sequences that anneal to the 3′ and 5′ ends of at least one of the parent polynucleotides under annealing conditions;
d) amplifying the fragments that anneal to each other to generate at least one polynucleotide sequence encoding one or more
protein motifs having altered characteristics as compared to the one or more protein motifs encoded by said parent polynucleotides.
|