Genetic Applications of an Inverse Polymerase Chain Reaction
TLDR
The feasibility of IPCR is shown by amplifying the sequences that flank an IS1 element in the genome of a natural isolate of Escherichia coli.Abstract:
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.read more
Citations
More filters
Patent
Modified nucleosides, nucleotides, and nucleic acids, and uses thereof
TL;DR: The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using them as discussed by the authors, as well as a detailed discussion of their properties.
Journal ArticleDOI
Exploitation of nucleic acid packaging signals to generate a novel influenza virus-based vector stably expressing two foreign genes.
Tokiko Watanabe,Shinji Watanabe,Takeshi Noda,Takeshi Noda,Yutaka Fujii,Yutaka Fujii,Yoshihiro Kawaoka,Yoshihiro Kawaoka +7 more
TL;DR: A virus whose HA and NA vRNA coding regions were replaced with those of vesicular stomatitis virus glycoprotein (VSVG) and green fluorescent protein (GFP) while retaining virion incorporation signals for these segments could serve as a model for the construction of influenza A viruses designed to express and/or deliver foreign genes.
Journal ArticleDOI
The Saccharomyces retrotransposon Ty5 integrates preferentially into regions of silent chromatin at the telomeres and mating loci.
TL;DR: Ty5 target preference extends the link between telomere structure and reverse transcription as carried out by telomerase and Drosophila retrotransposons and is predicted to be attributable to interactions between transposition intermediates and constituents of silent chromatin assembled at these sites.
Journal ArticleDOI
A versatile in vivo system for directed dissection of gene expression patterns
Daryl M. Gohl,Marion Silies,Xiaojing J. Gao,Sheetal Bhalerao,Francisco J. Luongo,Chun Chieh Lin,Christopher Potter,Thomas R. Clandinin +7 more
TL;DR: A convertible genetic platform, the integrase swappable in vivo targeting element (InSITE) system, that allows GAL4 to be replaced with any other sequence, placing different genetic effectors under the control of the same regulatory elements.
Journal ArticleDOI
Hierarchy among viral RNA (vRNA) segments in their role in vRNA incorporation into influenza A virions.
Yukiko Muramoto,Ayato Takada,Ken Fujii,Takeshi Noda,Kiyoko Iwatsuki-Horimoto,Shinji Watanabe,Taisuke Horimoto,Hiroshi Kida,Yoshihiro Kawaoka +8 more
TL;DR: A hierarchy among vRNA segments for virion incorporation is suggested and may imply intersegment association of vRNAs during virus assembly.
References
More filters
Journal ArticleDOI
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase
Randall Keichi Saiki,David H. Gelfand,Susanne Stoffel,Stephen J. Scharf,Russell Higuchi,Glenn Thomas Horn,Kary B. Mullis,Henry A. Erlich +7 more
TL;DR: A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction, which significantly improves the specificity, yield, sensitivity, and length of products that can be amplified.
Journal ArticleDOI
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.
Randall Keichi Saiki,Stephen J. Scharf,Fred A. Faloona,Kary B. Mullis,Glenn Thomas Horn,Henry A. Erlich,Norman Arnheim +6 more
TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
Book ChapterDOI
Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.
Kary B. Mullis,Fred A. Faloona +1 more
TL;DR: A method whereby a nucleic acid sequence can be exponentially amplified in vitro is described in the chapter, and the possibility of utilizing a heat-stable DNA polymerase is explored so as to avoid the need for addition of new enzyme after each cycle of thermal denaturation.
Journal ArticleDOI
Direct cloning and sequence analysis of enzymatically amplified genomic sequences
TL;DR: A method is described for directly cloning enzymatically amplified segments of genomic DNA into an M13 vector for sequence analysis and promises to be a rapid method for obtaining reliable genomic sequences from nanogram amounts of DNA.
Journal ArticleDOI
DNA typing from single hairs
TL;DR: Three different means of DNA typing are used for the determination of amplified DNA fragment length differences, hybridization with allele-specific oligonucleotide probes, and direct DNA sequencing on single human hairs to detect genetically variable mitochondrial and nuclear DNA sequences.