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Full-Text Articles in Life Sciences
Direct Genomic Sequencing Of Bacterial Dna: The Pyruvate Kinase I Gene Of Escherichia Coli, Osamu Ohara, Robert L. Dorit, Walter Gilbert
Direct Genomic Sequencing Of Bacterial Dna: The Pyruvate Kinase I Gene Of Escherichia Coli, Osamu Ohara, Robert L. Dorit, Walter Gilbert
Biological Sciences: Faculty Publications
The genomic sequencing procedure is applied to the direct sequencing of uncharacterized regions of bacterial DNA by a "multiplex walking" approach. Samples of bulk Escherichia coli DNA are cut with various restriction enzymes, subjected to chemical sequencing degradations, run in a sequencing gel, and transferred to nylon membranes. When a labeled oligomer is hybridized to a membrane, a sequence ladder appears wherever the probe lies near a restriction cut. New probes, based on sequence that lies beyond other restriction sites, are then synthesized, and the membranes are reprobed to reveal new sequence. Repeated cycles of oligomer probe synthesis and subsequent …
One-Sided Polymerase Chain Reaction: The Amplification Of Cdna, Osamu Ohara, Robert L. Dorit, Walter Gilbert
One-Sided Polymerase Chain Reaction: The Amplification Of Cdna, Osamu Ohara, Robert L. Dorit, Walter Gilbert
Biological Sciences: Faculty Publications
We report a rapid technique, based on the polymerase chain reaction (PCR), for the direct targeting, enhancement, and sequencing of previously uncharacterized cDNAs. This method is not limited to previously sequenced transcripts, since it requires only two adjacent or partially overlapping specific primers from only one side of the region to be amplified. These primers can be located anywhere within the message. The specific primers are used in conjunction with nonspecific primers targeted either to the poly(A)+ region of the message or to an enzymatically synthesized d( A) tail. Pairwise combinations of specific and general primers allow for the amplification …
A Field Study Using The Polymerase Chain Reaction (Pcr) To Screen For Brugia Microfilariae In Human And Animal Blood, Janet Glover, Steven A. Williams, Susanne Szabo, David Landry, Larry A. Mcreynolds, Taniawati Supali, Felix Partono
A Field Study Using The Polymerase Chain Reaction (Pcr) To Screen For Brugia Microfilariae In Human And Animal Blood, Janet Glover, Steven A. Williams, Susanne Szabo, David Landry, Larry A. Mcreynolds, Taniawati Supali, Felix Partono
Biological Sciences: Faculty Publications
Blood samples from 43 humans and 14 cats with Brugia microfilariae were analyzed in a field study in Tanjung Pinang, Indonesia. The study used the polymerase chain reaction (PCR) to compare the sensitivity of radioactive and biotinylated species-specific oligonuleotide probes. The cloning char- acterization of the Hha I repeat DNA family found in filarial parasites of the genus Brugia, and the development of species-specific probes for B.malayi and B.pahangi based on these repeats has been described elsewhere (PNAS USA 83: 797-801); Mol.Biochem. Parasitol. 28: 163-170). The use of radioisotopes for labelling DNA probes is both expensive and inconvenient. To replace …
Phase-Variation Of Pyelonephritis-Associated Pili In Escherichia Coli: Evidence For Transcriptional Regulation, L. B. Blyn, B. A. Braaten, C. A. White-Ziegler, D. H. Rolfson, D. A. Low
Phase-Variation Of Pyelonephritis-Associated Pili In Escherichia Coli: Evidence For Transcriptional Regulation, L. B. Blyn, B. A. Braaten, C. A. White-Ziegler, D. H. Rolfson, D. A. Low
Biological Sciences: Faculty Publications
The regulation of pyelonephritis-associated pill (pap) pflin gene transcription has been examined using two operons (pap-17 and pap-21) isolated from the pyelonephritogenic Escherichia coli strain C1212. DNA sequence analysis and E.coli minicell analysis were used to map two genes (papB and papl) within the pilin regulatory regions of both pap-17 and pap-21, and the protein products of these genes were identified. Pilin transcription, initiated at the papBA promoter, was monitored by constructing single copy operon fusions with lacZYA in E.coli K-12. Inocula- tion of E.coli (pap'-lac) strains onto solid M9 minimal medium containing glycerol and the Lac indicator X-gal (M9-Glycerol) …