Search results for "Restriction"
showing 10 items of 527 documents
Molecular diversity and growth features of Flavobacterium columnare strains isolated in Finland
2006
ABSTRACT: Columnaris disease caused by Flavobacterium columnare is a problem in fish farmingworldwide. During the last 15 yr, outbreaks have started to emerge in Finland. Flavobacteriumcolumnare Type Strain NCIMB 2248 T and 30 Finnish F. columnare isolates were studied usinganalysis of 16S rDNA by restriction-fragment length polymorphism (16S RFLP), length heterogeneityanalysis of polymerase chain reaction (LH-PCR) products, automated ribosomal intergenic spaceranalysis (ARISA), and 16S rDNA sequence analysis. All isolates fell into RFLP Genomovar I and hadthe same length in the LH-PCR analysis. Based on ARISA, 8 genetically different strains wereselected for further analyses. The growth of…
Assay for O6-alkylguanine-DNA-alkyltransferase using oligonucleotides containing O6-methylguanine in a BamHI recognition site as substrate
1992
Abstract Double-stranded oligonucleotides, 40 bases in length containing an O 6 -methylguanine in a Bam HI restriction site, were developed as substrates for the determination of human O 6 -alkylguanine-DNA-alkyltransferase (AGT). The assay proved highly sensitive and quantitative. After incubation of the 5′-end-labeled oligonucleotides with cell homogenates of peripheral blood lymphocytes, the DNA was digested with Bam HI. Cleavage with this restriction enzyme did not occur in the O 6 -methylguanine-containing oligonucleotide unless the fragment was repaired. The cleaved oligonucleotide was separated from the intact parent oligonucleotide by reverse-phase high-performance liquid chromatogr…
A new species of Pythium isolated from the Burgundy region in France.
1998
Pythium nodosum sp. nov. has been isolated from a soil sample taken in the Burgundy region in France. The fungus has spherical to variously shaped proliferating sporangia, smooth-walled oogonia which are crowded with different antheridial branches making a complicated knot around the former, and aplerotic oospores. Morphological and reproductive aspects of Pythium nodosum as well as the PCR of the internal transcribed spacer (ITS1) of the ribosomal nuclear DNA coupled with restriction fragment length polymorphism analysis are described here. The nucleotide sequences of ITS1 encoding 5.8S rRNA is also given.
Isolation of the DNA minisatellite probe MZ 1.3 and its application to DNA ‘fingerprinting’ analysis
1990
Abstract A minisatellite probe, MZ 1.3, detecting hypervariable fragment patterns was isolated from a human genomic library. A repetitive sequence of 27 bp length was identified which is contained in the probe approx. 40 times. The MZ 1.3 repeat shows variable homology of 53–73% to the repetitive sequence of the protein III gene of the bacteriophage M13 genome. Polymorphic restriction fragment patterns were found with MZ 1.3 using the enzymes Hinf I, BstN I, Hae III, Mbo I, PstI PvuII , and Rsa I. An average of 18 polymorphic fragments was observed using Hinf I as enzyme. The band sharing frequency after Hinf I digestion among unrelated individuals was determined to be 23.8 ± 7.2%. An examp…
Chimeric Genomes of Natural Hybrids of Saccharomyces cerevisiae and Saccharomyces kudriavzevii
2009
11 pages, 6 figures.-- PMID: 19251887 [PubMed].-- Printed version published Apr 2009.
A bicistronic vector backbone for rapid seamless cloning and chimerization of αβT-cell receptor sequences.
2020
To facilitate preclinical testing of T-cell receptors (TCRs) derived from tumor-reactive T-cell clones it is necessary to develop convenient and rapid cloning strategies for the generation of TCR expression constructs. Herein, we describe a pDONR™221 vector backbone allowing to generate Gateway™ compatible entry clones encoding optimized bicistronic αβTCR constructs. It harbors P2A-linked TCR constant regions and head-to-head-oriented recognition sites of the Type IIS restriction enzymes BsmBI and BsaI for seamless cloning of the TCRα and TCRβ V(D)J regions, respectively. Additional well-established TCR optimizations were incorporated to enhance TCR functionality. This included replacing of…
Emerging GII.4 norovirus variants affect children with diarrhea in Palermo, Italy in 2006
2008
Although the genetic/antigenic heterogeneity of human noroviruses (NoVs) is impressive, a few genogroup II strains of genotype 4 (GII.4) are dominant worldwide. GII.4 NoVs evolve rapidly and in the last 15 years six epidemic variants have been identified. In 2005-2006, surveillance of sporadic viral gastroenteritis in children in Palermo, Italy, resulted in the detection of NoV strains in 20.9% of the patients admitted to hospital. By restriction fragment length polymorphism (RFLP) and sequence analysis of region A in the RNA-dependent RNA-polymerase (RdRp) gene, 59 NoV strains were successfully characterized. Eighty-one percent of the strains were characterized as GII.4, 14% as GIIb/Hilver…
Polyphasic identification of yeasts isolated from bark of cork oak during the manufacturing process of cork stoppers.
2003
A two-step protocol was used for the identification of 52 yeasts isolated from bark of cork oak at initial stages of the manufacturing process of cork stoppers. The first step in the identification was the separation of the isolates into groups by their physiological properties and RFLPs of the ITS-5.8S rRNA gene. The second step was the sequencing of the D1/D2 domains of the 26S rRNA gene of selected isolates representing the different groups. The results revealed a predominance of basidiomycetous yeasts (11 species), while only two species represented the ascomycetous yeasts. Among the basidiomycetous yeasts, members representing the species Rhodosporidium kratochvilovae and Rhodotorula n…
Molecular analysis of the nitrate-reducing community from unplanted and maize-planted soils
2002
ABSTRACT Microorganisms that use nitrate as an alternative terminal electron acceptor play an important role in the global nitrogen cycle. The diversity of the nitrate-reducing community in soil and the influence of the maize roots on the structure of this community were studied. The narG gene encoding the membrane bound nitrate reductase was selected as a functional marker for the nitrate-reducing community. The use of narG is of special interest because the phylogeny of the narG gene closely reflects the 16S ribosomal DNA phylogeny. Therefore, targeting the narG gene provided for the first time a unique insight into the taxonomic composition of the nitrate-reducing community in planted an…
Natural hybrids fromSaccharomyces cerevisiae,Saccharomyces bayanusandSaccharomyces kudriavzeviiin wine fermentations
2006
Several wine isolates of Saccharomyces were analysed for six molecular markers, five nuclear and one mitochondrial, and new natural interspecific hybrids were identified. The molecular characterization of these Saccharomyces hybrids was performed based on the restriction analysis of five nuclear genes ( CAT8 , CYR1 , GSY1 , MET6 and OPY1 , located in different chromosomes), the ribosomal region encompassing the 5.8S rRNA gene and the two internal transcribed spacers, and sequence analysis of the mitochondrial gene COX2 . This method allowed us to identify and characterize new hybrids between Saccharomyces cerevisiae and Saccharomyces kudriavzevii , between S. cerevisiae and Saccharomyces ba…