Overlap Extension PCR: An Efficient Method for Transgene Construction

Methods in Molecular Biology

Volumn 772, Issue , 2011, Pages 459-470

  Nelson, Matthew D ^a   Fitch, David H A ^a  

^a NEW YORK UNIVERSITY   (United States)

Author keywords

Cloning; Gene expression; Hybrid genes; PCR; Site directed mutagenesis; Transgenics

Indexed keywords

OLIGONUCLEOTIDE;

AGAR GEL ELECTROPHORESIS; ANIMAL; ARTICLE; CAENORHABDITIS ELEGANS; GENETICS; METABOLISM; METHODOLOGY; POLYMERASE CHAIN REACTION; SITE DIRECTED MUTAGENESIS; TRANSGENE;

ANIMALS; CAENORHABDITIS ELEGANS; ELECTROPHORESIS, AGAR GEL; MUTAGENESIS, SITE-DIRECTED; OLIGONUCLEOTIDES; POLYMERASE CHAIN REACTION; TRANSGENES;

EID: 84857206161     PISSN: 10643745     EISSN: 19406029     Source Type: Book Series    
DOI: 10.1007/978-1-61779-228-1_27     Document Type: Chapter

References (12)

1. Chalfie M, Tu Y, Euskirchen G et al (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805
2. Crocker J, Tamori Y, Erives A (2008) Evolution acts on enhancer organization to fine-tune gradient threshold readouts. PLoS Biol 6:e263
3. Hamamatsu N, Aita T, Nomiya Y et al (2005) Biased mutation-assembling: an efficient method for rapid directed evolution through simultaneous mutation accumulation. Protein Eng Des Sel 18:265–271
4. Mullis K, Faloona F, Scharf S et al (1986) Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harb Symp Quant Biol 51:263–273
5. Horton RM, Hunt HD, Ho SN et al (1989) Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene 77:61–68
6. Yon J, Fried M (1989) Precise gene fusion by PCR. Nucleic Acids Res 17:4895
7. Yolov AA, Shabarova ZA (1990) Constructing DNA by polymerase recombination. Nucleic Acids Res 18:3983–3986
8. Lundberg KS, Shoemaker DD, Adams MW et al (1991) High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus. Gene 108:1–6
9. Shevchuk NA, Bryksin AV, Nusinovich YA et al (2004) Construction of long DNA molecules using long PCR-based fusion of several fragments simultaneously. Nucleic Acids Res 32:e19
10. Heckman KL, Pease LR (2007) Gene splicing and mutagenesis by PCR-driven overlap extension. Nat Protoc 2:924–932
11. Ho SN, Hunt HD, Horton RM et al (1989) Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59
12. Fire A, Harrison SW, Dixon D (1990) A modular set of lacZ fusion vectors for studying gene expression in Caenorhabditis elegans. Gene 93:189–198