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
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Cloning and characterization of the genes for biosynthesis of the compatible solute ectoine in the moderately halophilic bacterium Halobacillus dabanensis D-8(T).
TL;DR: A 11.2-kb fragment containing the ectABC genes of the biosynthetic pathway of ectoine from the Gram-positive, moderately halophilic bacterium Halobacillus dabanensis D-8T was obtained by inverse polymerase chain reaction and the amino-acid sequence deduced was highly homologous that from Virgibacillus pantethenticus.
Journal ArticleDOI
Isolation and sequence analysis of a β-tubulin gene from arbuscular mycorrhizal fungi
Zola Msiska,Joseph B. Morton +1 more
TL;DR: A full-length β-tubulin gene has been cloned and sequenced from Gigaspora gigantea and Glomus clarum, two arbuscular mycorrhizal fungi (AMF) species in the phylum Glomeromyota, and the amino acid sequence is most similar to that of fungi in the Chytridiomycota.
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Chloramphenicol resistance transposable element TnSs1 of Streptococcus suis, a transposon flanked by IS6-family elements.
TL;DR: A new transposon, designated TnSs1, which contains a chloramphenicol acetyltransferase gene flanked by direct repeats of an IS6-family element was found in a field isolate of Streptococcus suis and showed cointegrate between the S. suis genome and the vector.
Journal ArticleDOI
Alcohol induced silencing of gibberellin 20-oxidases in Kalanchoe blossfeldiana
TL;DR: Reduction in the amounts of active gibberellic acids in elongating cuttings from the ornamental crop Kalanchoe blossfeldiana were pursued by genetic manipulation as an alternative to synthetic growth regulators to indicate that optimisation of the ethanol treatments can enable us to produce more compact growing plants still maintaining normal flowering.
Journal ArticleDOI
Analysis of a gene (vch) encoding hemolysin isolated and sequenced from Vibrio campbellii
TL;DR: This study describes the amplification, localization, and sequence analysis of a hemolysin gene from type strain V. campbellii NBRC 15631--the first report of a full-length hemoly sin gene for the species.
References
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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.
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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.