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
Journal ArticleDOI
The role of CBF transcriptional activators in two Citrus species (Poncirus and Citrus) with contrasting levels of freezing tolerance
TL;DR: This study provides the first evidence of a correlation between CBF expression (timing and quantity) and the degree of cold tolerance in two closely related species with wide differences in cold tolerance.
Journal ArticleDOI
Barbiturase, a Novel Zinc-containing Amidohydrolase Involved in Oxidative Pyrimidine Metabolism
TL;DR: The characteristics and gene organization of barbiturase suggested that it is a novel zinc-containing amidhydrolase that should be grouped into a new family of the amidohydrolases superfamily.
Journal ArticleDOI
Comparative structure and characterization of a CDKN2 gene in a Xiphophorus fish melanoma model
Steven Kazianis,Donald C. Morizot,L Della Coletta,Dennis A. Johnston,Bruce Woolcock,J.R. Vielkind,Rodney S. Nairn +6 more
TL;DR: Comparative sequence analysis suggests that fish CDKN2X is similarly related to all four mammalian gene family members, and may represent a descendant of an ancestral prototypic CD KN2 gene.
Journal ArticleDOI
Expression of a doublesex homologue is altered in sexual mosaics of Ostrinia scapulalis moths infected with Wolbachia
TL;DR: RT-PCR analysis indicated that Osdsx was transcribed in a sex-specific manner in all somatic tissues examined, regardless of developmental stage, and provides the first evidence that Wolbachia manipulates the sex of its host by interfering either with the sex- specific splicing of OsDSx itself or with another upstream sex determination process.
Journal ArticleDOI
The Aspergillus nidulans GATA transcription factor gene areB encodes at least three proteins and features three classes of mutation.
Helen Conlon,Ivo Zadra,Hubertus Haas,Herbert N. Arst,Herbert N. Arst,Meriel G. Jones,Mark X. Caddick +6 more
TL;DR: Three classes of mutation are characterized in areB; the first are loss‐of‐function mutations that terminate the polypeptides within or before the GATA domain, and two novel gene fusions transform the putative negative‐acting transcription factor into an activator that can partially replace areA.
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.