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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flagellar genes in Campylobacter jejuni is associated with formation of the flagellar secretory apparatus
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Construction and characterization of Streptococcus suis-Escherichia coli shuttle cloning vectors.
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Membranous Cells in Nasal-Associated Lymphoid Tissue: A Portal of Entry for the Respiratory Mucosal Pathogen Group A Streptococcus
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Circadian clock protein KaiC forms ATP-dependent hexameric rings and binds DNA
Tetsuya Mori,Sergei V. Saveliev,Yao Xu,Walter F. Stafford,Michael M. Cox,Ross B. Inman,Carl Hirschie Johnson +6 more
TL;DR: It is shown here that KaiC also forms ring complexes with a central pore that can be visualized by electron microscopy, and this data support the inclusion of KaiC into the RecA/DnaB superfamily and have important implications for enzymatic activity ofkaiC in the circadian clock mechanism that regulates global changes in gene expression patterns.
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Phage conversion of exfoliative toxin A production in Staphylococcus aureus.
Takayuki Yamaguchi,Tetsuya Hayashi,Hideto Takami,Kaoru Nakasone,Makoto Ohnishi,Keisuke Nakayama,Sakuo Yamada,Hitoshi Komatsuzawa,Motoyuki Sugai +8 more
TL;DR: A temperate phage that encodes ETA is isolated and the complete nucleotide sequence of the φETA genome is determined, which suggests that the eta gene is acquired by horizontal gene transfer in S. aureus.
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.
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.