Genetic Applications of an Inverse Polymerase Chain Reaction

INVESTIGATIONS

Genetic Applications of an Inverse Polymerase Chain Reaction

H. Ochman, A. S. Gerber and D. L. Hartl
Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110

A method is presented for the rapid in vitro amplification of DNA sequences that flank a region of known sequence. The method uses the polymerase chain reaction (PCR), but it has the primers oriented in the reverse direction of the usual orientation. The template for the reverse primers is a restriction fragment that has been ligated upon itself to form a circle. This procedure of inverse PCR (IPCR) has many applications in molecular genetics, for example, the amplification and identification of sequences flanking transposable elements. In this paper we show the feasibility of IPCR by amplifying the sequences that flank an IS1 element in the genome of a natural isolate of Escherichia coli.

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				J. Zupan, C. A. Hackworth, J. Aguilar, D. Ward, and P. Zambryski *VirB1 Promotes T-Pilus Formation in the vir-Type IV Secretion System of Agrobacterium tumefaciens** J. Bacteriol., September 15, 2007; 189(18): 6551 - 6563. [Abstract] [Full Text] [PDF]

				S. Wang, J. He, Z. Cui, and S. Li Self-Formed Adaptor PCR: a Simple and Efficient Method for Chromosome Walking Appl. Envir. Microbiol., August 1, 2007; 73(15): 5048 - 5051. [Abstract] [Full Text] [PDF]

				H. Wang, M. Johnston, and R. D. Mitra Calling cards for DNA-binding proteins Genome Res., August 1, 2007; 17(8): 1202 - 1209. [Abstract] [Full Text] [PDF]

				I. Mruk and T. Kaczorowski A Rapid and Efficient Method for Cloning Genes of Type II Restriction-Modification Systems by Use of a Killer Plasmid Appl. Envir. Microbiol., July 1, 2007; 73(13): 4286 - 4293. [Abstract] [Full Text] [PDF]

				A. C. Silver, Y. Kikuchi, A. A. Fadl, J. Sha, A. K. Chopra, and J. Graf Interaction between innate immune cells and a bacterial type III secretion system in mutualistic and pathogenic associations PNAS, May 29, 2007; 104(22): 9481 - 9486. [Abstract] [Full Text] [PDF]

				D. V. Volokhov, S. Duperrier, A. A. Neverov, J. George, C. Buchrieser, and A. D. Hitchins The Presence of the Internalin Gene in Natural Atypically Hemolytic Listeria innocua Strains Suggests Descent from L. monocytogenes Appl. Envir. Microbiol., March 15, 2007; 73(6): 1928 - 1939. [Abstract] [Full Text] [PDF]

				S. Goh, P. F. Ong, K. P. Song, T. V. Riley, and B. J. Chang The complete genome sequence of Clostridium difficile phage {phi}C2 and comparisons to {phi}CD119 and inducible prophages of CD630 Microbiology, March 1, 2007; 153(3): 676 - 685. [Abstract] [Full Text] [PDF]

				S. M. Alavi, S. Poussier, and C. Manceau Characterization of ISXax1, a Novel Insertion Sequence Restricted to Xanthomonas axonopodis pv. phaseoli (Variants fuscans and non-fuscans) and Xanthomonas axonopodis pv. vesicatoria Appl. Envir. Microbiol., March 1, 2007; 73(5): 1678 - 1682. [Abstract] [Full Text] [PDF]

				M. Annaheim and B. Lanzrein Genome organization of the Chelonus inanitus polydnavirus: excision sites, spacers and abundance of proviral and excised segments J. Gen. Virol., February 1, 2007; 88(2): 450 - 457. [Abstract] [Full Text] [PDF]

				T. Kotani, H. Yurimoto, N. Kato, and Y. Sakai Novel Acetone Metabolism in a Propane-Utilizing Bacterium, Gordonia sp. Strain TY-5 J. Bacteriol., February 1, 2007; 189(3): 886 - 893. [Abstract] [Full Text] [PDF]

				S. Shulami, G. Zaide, G. Zolotnitsky, Y. Langut, G. Feld, A. L. Sonenshein, and Y. Shoham A Two-Component System Regulates the Expression of an ABC Transporter for Xylo-Oligosaccharides in Geobacillus stearothermophilus Appl. Envir. Microbiol., February 1, 2007; 73(3): 874 - 884. [Abstract] [Full Text] [PDF]

				M. Ozawa, K. Fujii, Y. Muramoto, S. Yamada, S. Yamayoshi, A. Takada, H. Goto, T. Horimoto, and Y. Kawaoka Contributions of Two Nuclear Localization Signals of Influenza A Virus Nucleoprotein to Viral Replication J. Virol., January 1, 2007; 81(1): 30 - 41. [Abstract] [Full Text] [PDF]

				W. An, J. S. Han, S. J. Wheelan, E. S. Davis, C. E. Coombes, P. Ye, C. Triplett, and J. D. Boeke Active retrotransposition by a synthetic L1 element in mice PNAS, December 5, 2006; 103(49): 18662 - 18667. [Abstract] [Full Text] [PDF]