Skip to Content
Find More Like This
Return to Search

Recombination of polynucleotide sequences using random or defined primers

United States Patent

November 28, 2000
View the Complete Patent at the US Patent & Trademark Office
A method for in vitro mutagenesis and recombination of polynucleotide sequences based on polymerase-catalyzed extension of primer oligonucleotides is disclosed. The method involves priming template polynucleotide(s) with random-sequences or defined-sequence primers to generate a pool of short DNA fragments with a low level of point mutations. The DNA fragments are subjected to denaturization followed by annealing and further enzyme-catalyzed DNA polymerization. This procedure is repeated a sufficient number of times to produce full-length genes which comprise mutants of the original template polynucleotides. These genes can be further amplified by the polymerase chain reaction and cloned into a vector for expression of the encoded proteins.
Arnold; Frances H. (Pasadena, CA), Shao; Zhixin (Pasadena, CA), Affholter; Joseph A. (Midland, MI), Zhao; Huimin H (San Diego, CA), Giver; Lorraine J. (Sunnyvale, CA)
California Institute of Technology (Pasadena, CA)
08/ 905,359
August 4, 1997
The U.S. Government has certain rights in this invention pursuant to Grant No. DE-FG02-93-CH10578 awarded by the Department of Energy and Grant No. N00014-96-1-0340 awarded by the Office of Naval Research.