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Open AccessJournal ArticleDOI

Genetic Applications of an Inverse Polymerase Chain Reaction

Howard Ochman, +2 more
- 01 Nov 1988 - 
- Vol. 120, Iss: 3, pp 621-623
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.

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Journal ArticleDOI

Sodium selenite induces apoptosis in cultured cortical neurons with special concomitant changes in expression of the apoptosis-related genes.

TL;DR: It is suggested that the sodium selenite is effective for inducing apoptosis in cultured cortical neurons and that relevant changes in expression of several apoptosis-related genes might further the understanding of the mechanism(s) that initiates and maintains the apoptotic processes.
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Cloning and characterization of a NAD+-dependent glycerol-3-phosphate dehydrogenase gene from Candida glycerinogenes, an industrial glycerol producer.

TL;DR: Cloned and characterized a 4900-bp genomic fragment containing the CgGPD gene encoding a glycerol-3-phosphate dehydrogenase homologous to GPD genes in other yeasts using degenerate primers in conjunction with inverse PCR, suggesting that it is a functional GPD protein.
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The ribulose monophosphate pathway operon encoding formaldehyde fixation in a thermotolerant methylotroph, Bacillus brevis S1.

TL;DR: Enzyme induction and Northern blot analyses revealed that both the hps and phi genes are induced by methanol or ethanol, and that their expression is controlled polycistronically at the transcription stage.
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A forward genetic approach in Chlamydomonas reinhardtii as a strategy for exploring starch catabolism.

TL;DR: A screen to study the mobilization of starch in the unicellular green alga Chlamydomonas reinhardtii relies on starch synthesis accumulation during nitrogen starvation followed by the supply of nitrogen and the switch to darkness, and characterizes the nature of the genes disrupted in the selected mutants.
Journal ArticleDOI

A mutant screen reveals RNase E as a silencer of group II intron retromobility in Escherichia coli

TL;DR: The results suggest that RNase E, which is the central component of the RNA degradosome, could regulate retrohoming levels in response to cellular physiology.
References
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Journal ArticleDOI

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase

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.

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.

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.
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