Circular site-directed mutagenesis

Details Circular site-directed mutagenesis Circular site-directed mutagenesis

2006-11-07

2006-11-07

Horlick, Kenneth R. (Department: 1637)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

C435S091200

Reexamination Certificate

active

07132265

ABSTRACT:
The invention provides improved methods of introducing site-directed mutations into circular DNA molecules of interest by means of mutagenic primer pairs. The mutagenic primer pairs are also selected so as to be either completely complementary or partially complementary to each other, wherein the mutation site (or sites) is located within the region of complementarity. A mutagenic primer pair is annealed to opposite strands of a circular DNA molecule containing the DNA sequence to be mutagenized. After annealing, first and second mutagenized DNA strands, each incorporating a member of the mutagenic oligonucleotide primer pair is synthesized by a linear cyclic amplification reaction. After the linear cyclic amplification mediated synthesis step is completed, the reaction mixture is treated with a selection enzyme that digests the parental template strands. After the digesting step, a double-stranded circular DNA intermediate is formed. The double-stranded circular DNA intermediates is transformed in suitable competent host cells and closed circular double-stranded DNA corresponding to the parental template molecules, but containing the desired mutation or mutations of interest, may be conveniently recovered from the transformed cells. The invention also provide kits for site-directed mutagenesis in accordance with methods of the present invention.

REFERENCES:
patent: 5286632 (1994-02-01), Jones
patent: 5436149 (1995-07-01), Barnes
patent: 5512463 (1996-04-01), Stemmer
patent: 5514568 (1996-05-01), Stemmer
patent: 5789166 (1998-08-01), Bauer et al.
patent: 5932419 (1999-08-01), Bauer et al.
patent: 6391548 (2002-05-01), Bauer et al.
patent: 6713285 (2004-03-01), Bauer et al.
patent: WO 93/12257 (1993-06-01), None
patent: WO 93/18175 (1993-09-01), None
U.S. Appl. No. 10/811,062, filed Mar. 25, 2004.
Preliminary Amendment dated Mar. 25, 2004, in U.S. Appl. No. 10/811,062, filed Mar. 25, 2004.
Barnes,Proc. Natl. Acad. Sci. U.S.A.,91:2216-2220 (1994).
Cheng et al.,Proc. Natl. Acad. Sci. U.S.A.,91:5695-5699 (1994).
Deng et al.,Analytical Biochemistry,200:81-88 (1992).
Drusta et al.,Nucleic Acids Research,20:922 (1991).
Hultman et al.,Nucleic Acids Research,18:5107-5112 (1990).
Jones et al., Methods: A Companion to Methods in Enzymology, 2(1):2-10 (1991).
Kunkel,Proc. Natl. Acad. Sci. U.S.A.,82:488-492 (1985).
Lai et al.,Nucleic Acids Research,21:3977-3980 (1993).
McClary et al., Biotechniques, 7(3):282-287 (1989).
Stemmer and Morris,BioTechniques,13:214-220 (1992).
Stemmer et al.,BioTechniques,14:256-265 (1993).
Taylor et al.,Nucleic Acids Research,13:8765-8785 (1985).
Vilardaga et al.,BioTechniques,18:604-606 (1995).
Watkins et al.,BioTechniques,5:700-704 (1993).
Weiner et al.,Gene,126:35-41 (1993).
Weiner et al.,PCR Methods&Applications,4(3):s131-s136 (1994).
Weiner et al.,Gene,151(1):119-123 (1994).
Yao et al.,PCR Methods Appl.,1:205-207 (1992).
Supplementary European Search Report, dated Aug. 19, 2002.
Rashtchian et al.,PCR Methods and Applications,2:124-130 (1992).
Office Action in U.S. Appl. No. 08/567,881, mailed on Oct. 17, 1996.
Notice of Allowability, Examiner's Amendment, and Examiner Interview Summary Record in U.S. Appl. No. 08/567,881, mailed on Apr. 16, 1997.
Office Action in U.S. Appl. No. 08/844,524, mailed on Oct. 15, 1997.
Response filed in U.S. Appl. No. 08/844,524, on Apr. 15, 1998.
Office Action in U.S. Appl. No. 08/844,524, mailed on Jul. 17, 1998.
Response filed in U.S. Appl. No. 08/844,524, on Dec. 10, 1998.
Notice of Allowability and Interview Summary in U.S. Appl. No. 08/844,524, on Jan. 22, 1999.
Office Action in U.S. Appl. No. 09/274,383, mailed on Aug. 16, 2000.
Response filed in U.S. Appl. No. 09/274,383, on Jan. 16, 2001.
Examiner's Amendment in U.S. Appl. No. 09/274,383, mailed on Jun. 29, 2001.
Response filed in U.S. Appl. No. 09/274,383, on Jun. 29, 2001.
Preliminary Amendment filed in U.S. Appl. No. 10/144,652, on Aug. 16, 2002.
Office Action in U.S. Appl. No. 10/144,652, mailed on Dec. 13, 2002.
Response filed in U.S. Appl. No. 10/144,652, on Mar. 11, 2003.
Office Action in U.S. Appl. No. 10/144,652, mailed on May 30, 2003.
Response filed in U.S. Appl. No. 10/144,652, on Sep. 3, 2003.
Office Action in U.S. Appl. No. 10/811,062, mailed on Aug. 15, 2005.
Response filed in U.S. Appl. No. 10/811,062, on Nov. 10, 2005.
Office Action in U.S. Appl. No. 10/811,062, mailed on Jan. 26, 2006.
International Search Report for International Patent Application No. PCT/US96/19387, mailed on Mar. 21, 1997.
PCT Written Opinion for International Patent Application No. PCT/US96/19387, mailed on Dec. 10, 1997.
PCT International Preliminary Examination Report for International Patent Application No. PCT/US96/19387, dated Feb. 18, 1998.
European Patent Office Examination Report for European Patent Application No. 96942905, dated May 13, 2003.
European Patent Office Examination Report for European Patent Application No. 96942905, dated Jul. 2, 2004.
Official Action in Japanese Patent Application No. JP 9-521433, mailed on Oct. 21, 2003.
English Translation of Official Action in Japanese Patent Application No. JP 9-521433, mailed on Oct. 21, 2003, provided by A. Aoki, Ishida & Associates, Tokyo, Japan.
Official Action in Japanese Patent Application No. JP 9-521433, mailed on Apr. 19, 2005.
English Translation of Official Action in Japanese Patent Application No. JP 9-521433, mailed on Apr. 19, 2005, provided by A. Aoki, Ishida & Associates, Tokyo, Japan.
Official Action in Canadian Patent Application No. 2,239,879, mailed on Mar. 9, 2005.
Response filed in U.S. Appl. No. 10/811,062, on Jun. 9, 2006.

Affiliated with

Also associated with

Rating

Circular site-directed mutagenesis does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Circular site-directed mutagenesis, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Circular site-directed mutagenesis will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3700982