Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction.

Gene Splicing by Overlap Extension or "gene SOEing" is a PCR-based method of recombining DNA sequences without reliance on restriction sites and of directly generating mutated DNA fragments in vitro. By modifying the sequences incorporated into the 5'-ends of the primers, any pair of polymerase chain reaction products can be made to share a common sequence at one end. Under polymerase chain reaction conditions, the common sequence allows strands from two different fragments to hybridize to one another, forming an overlap. Extension of this overlap by DNA polymerase yields a recombinant molecule. This powerful and technically simple approach offers many advantages over conventional approaches for manipulating gene sequences.

[1]  V. E. Williams,et al.  Regions of allelic hypervariability in the murine Aα immune response gene , 1983, Cell.

[2]  K. Mullis,et al.  Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. , 1986, Cold Spring Harbor symposia on quantitative biology.

[3]  H. Mcdevitt,et al.  Sequence analysis and structure-function correlations of murine q, k, u, s, and f haplotype I-A beta cDNA clones. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[4]  K. Mullis,et al.  Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. , 1987, Methods in enzymology.

[5]  L. Leinwand,et al.  Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. , 1988, BioTechniques.

[6]  R. Saiki,et al.  A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. , 1988, Nucleic acids research.

[7]  T. Kadowaki,et al.  Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis. , 1989, Gene.

[8]  M. Aigle,et al.  Directed mutagenesis using PCR. , 1989, Nucleic acids research.

[9]  S. Ho,et al.  Site-directed mutagenesis by overlap extension using the polymerase chain reaction. , 1989, Gene.

[10]  S. Pääbo,et al.  Ancient DNA and the polymerase chain reaction. The emerging field of molecular archaeology. , 1989, The Journal of biological chemistry.

[11]  S. Ho,et al.  Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. , 1989, Gene.

[12]  J. Yon,et al.  Precise gene fusion by PCR. , 1989, Nucleic acids research.

[13]  H. Hunt,et al.  The functional significance of two amino acid polymorphisms in the antigen-presenting domain of class I MHC molecules. Molecular dissection of Kbm3. , 1989, Journal of immunology.

[14]  G. Sarkar,et al.  The "megaprimer" method of site-directed mutagenesis. , 1990, BioTechniques.