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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Strategies for cloning unknown cellular flanking DNA sequences from foreign integrants.
TL;DR: Techniques, including inverse polymerase chain reaction (IPCR), novel Alu-PCR and vectorette- or splinkerette- PCR are introduced as possible strategies for cloning flanking DNA regions of the integrants.
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Analysis of the human GDNF gene reveals an inducible promoter, three exons, a triplet repeat within the 3′-UTR and alternative splice products
Lena Grimm,Elke Holinski-Feder,Jens Teodoridis,Beatrix Scheffer,Dirk Schindelhauer,Thomas Meitinger,Marius Ueffing +6 more
TL;DR: The genomic structure and characterization of the promoter of the human GDNF gene shows that the hGDNF gene is driven by a TATA-containing promoter preceding exon 1, and fibroblast growth factor 2, tetradecanoyl 12-phorbol acetate, an inflammatory agent, and cAMP increase promoter activity, suggesting that GDNF transcriptional regulation is a target of exogenous signals.
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P element homing to the Drosophila bithorax complex.
Welcome Bender,A. Hudson +1 more
TL;DR: P elements containing a 7 kb DNA fragment from the middle of the Drosophila bithorax complex insert preferentially into the bith orax complex or into the adjacent chromosome regions, similar to that reported for the engraved promoter.
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Transcriptional activation of Cor/Lea genes and increase in abiotic stress tolerance through expression of a wheat DREB2 homolog in transgenic tobacco
TL;DR: Results clearly indicate that WDREB2 acts as a transcription factor and positively regulates Wdhn13, Wrab17, Wrabs18, and Wrab19 in the development of multiple abiotic stress tolerance in wheat.
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Molecular characterization of the S locus in two self-incompatible Brassica napus lines.
TL;DR: The chromosomal region carrying the SLG and SRK genes has been studied and two novel S locus genes are designated, SLL1 and SLL2, which are not clear whether these genes function in self-incompatibility or serve some other cellular roles in pollen-pistil functions.
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.