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Results for “Polymorphism, Restriction Fragment Length”

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At least 937 records · Page 52Linked to original sources
PubMed2004

[Study on clinical isolates of human cytomegalovirus found in Urumqi by restriction fragment length polymorphism of PCR products].

BACKGROUND: The study was designed to investigate the status of molecular epidemiology of HCMV in Urumqi through genetic comparison of clinical isolates. METHODS: DNA sequences of 2.0-2.6 kb were amplified by polymerase chain reaction from three relatively conservative gene regions (DNA polymerase, glycoproteins H, and major immediate-early antigen) of 28 clinical HCMV strains and then were analysed by restriction enzymes. RESULTS: The restriction patterns of the clinical isolates which did not have relation in epidemiology were greatly different, but the patterns of the clinical isolates related in epidemiology such as strains paired in mother and infant were quite similar. Of eight mother and infant pairs, from whom HCMV were isolated, four pairs showed identity of restriction profiles within each pair for all three amplified regions, four pairs showed differences between mother and infant. CONCLUSION: These results confirm the high degree of genetic variability among cytomegalovirus strains in Urumqi. Analysis of PCR-RFLP can indicate transmission of HCMV infection and facilitate its molecular epidemiologic studies.

China↗
PubMed1990

Restriction fragment length polymorphism for the Yc subunit gene of rat liver glutathione S-transferase.

An altered expression of the Yc subunit gene of rat glutathione S-transferase (GST) in the liver of the LEC rat, which is a mutant strain with spontaneous hereditary hepatitis associated with severe jaundice, has been reported. To provide further information concerning the structure of the Yc subunit gene, we carried out the Southern blot hybridization analysis of DNA samples from rats of eight different inbred strains including LEC with cDNA complementary to mRNA specific for the Yc subunit of rat liver GST as a probe. The hybridization patterns of the DNA samples from rats belonging to the different inbred strains showed interstrain variation in the length of restriction fragments with four restriction endonucleases. Since the DNA samples prepared from several rats of one inbred strain gave an identical hybridization pattern, the restriction fragment patterns for the Yc gene could be used as markers for genetic monitoring of inbred rat strains. Although the altered expression of Yc-Yc activity of GST has been observed in the liver of the LEC rat, the characteristic changes in the gene structure of the Yc subunit of LEC rat were not detected in the present hybridization analysis.

Animals↗
PubMed1993

A sensitive restriction fragment length polymorphism method to detect CAA-->AAA mutations at codon 61 of Ha-ras.

A rapid and sensitive assay was developed to detect CAA-->AAA mutations at codon 61 of Ha-ras. The region surrounding codon 61 was amplified by the polymerase chain reaction (PCR) using one primer containing a mismatch at the second position of codon 60. Using this primer creates an Msel restriction enzyme site if codon 61 carries the C.G-->A.T transversion. An aliquot of the second PCR primer was 5'-end-labeled with 32P to increase the sensitivity of detection of the PCR product. After cleavage with Msel, DNA was electrophoresed on a nondenaturing polyacrylamide gel, and the products were visualized by autoradiography. The sensitivity of this assay was such that the mutation could be detected when present in only one of 200 alleles. DNA samples from spontaneous Crl:CD-1(ICR)BR mouse liver tumors were analyzed using this method. Nine of 38 samples contained the mutation, and in one of those nine, the mutation had not been previously detected by either direct sequencing of tumor DNA or by sequencing the DNA from NIH 3T3 cells transfected with the tumor DNA.

3T3 Cells↗
PubMed1991

Characterization of highly virulent Escherichia coli strains by ribosomal DNA restriction fragment length polymorphism.

Patterns of ribosomal DNA polymorphism were examined to compare carboxylesterase B type B1 strains and B2 strains of Escherichia coli isolated from extra-intestinal infections. DNA from 14 type B2 strains showing the presence of alpha-haemolysin and mannose-resistant haemagglutinin and lethality to mice and 14 type B1 strains lacking these characteristics, was digested with HindIII, EcoRI, BamHI or BglII restriction enzymes and analysed by Southern blotting. The obtained ribotypes clearly differentiated the B2 strains from the B1 strains. These results indicate that genotypes of the highly virulent B2 strains are different from that of the less virulent B1 strains.

Blotting, Southern↗
PubMed2003

Seasonal population changes in the restriction fragment length polymorphism (RFLP) patterns from PCR-amplified 16S rRNA genes of predominant ribotypes in microbial mat samples from the Ebro Delta (Spain).

The stratified benthic microbial communities, which developed as a consequence of the physico-chemical gradients and the physiology of the inhabiting microorganisms, from Ebro Delta microbial mats were analyzed. 16S rRNA eubacterial and archaeal genes were amplified by polymerase chain reaction (PCR). PCR products were separately digested with three different restriction enzymes (AluI, HinfI, and RsaI) and later separated by horizontal electrophoresis to determine whether changes of predominant ribotypes are occurring over a period of a year. Comparison of total restriction patterns was performed by scoring similarities by using the Jaccard coefficient and then building a multidimensional scaling (MDS) map from the resulting similarities matrix. The three enzymes gave a consistent result, a seasonal distribution instead of a spatial and/or physiological one.

Archaea↗
PubMed2002

Simultaneous genotyping of single nucleotide polymorphisms in the IL-1 gene complex by multiplex polymerase chain reaction-restriction fragment length polymorphism.

The interleukin-1 (IL-1) gene complex consists of the IL-1alpha, IL-1beta and IL-1 receptor antagonist genes. Single-nucleotide polymorphisms (SNP) in all three genes have been associated with human diseases. In this study, primers containing mismatches at 1-3 nucleotide positions were designed to incorporate a restriction site for endonuclease AlwNI or XcmI in the presence of allele-specific nucleotides at the polymorphic positions. Based on this technique, a simple and robust multiplex polymerase chain reaction/restriction fragment length polymorphism (multiplex PCR/RFLP) assay was developed to determine simultaneously three to four informative SNPs (IL-1beta/+3954, IL-1beta/-511 and IL-1Ra/9261 or IL-1alpha/-889, IL-1beta/-31, IL-1beta/5810 and IL-1Ra/11100 SNPs) in the IL-1 gene complex.

Alleles↗
PubMed1997

Three different GB virus C/hepatitis G virus genotypes. Phylogenetic analysis and a genotyping assay based on restriction fragment length polymorphism.

The 5'-untranslated region (5'-UTR) sequences of 33 GB virus C/hepatitis G virus (GBV-C/HGV) obtained from different geographic areas were determined through reverse-transcription polymerase chain reaction and dideoxy chain termination sequencing, the alignment of sequences, the estimation of the number of nucleotide substitution per site, and construction of phylogenetic trees. The 5'-UTR of GBV-HGV was found to be heterogeneous, with 70.9-99.5% homology. Three distinct phylogenetic branches were observed consistently in all phylogenetic trees. GBV-C is the prototype for one, HGV for another, and there is a new branch which consisted of GBV-C/HGV isolates from Asia. Genotype-specific restriction sites for the restriction enzymes, ScrFI and BsmFI, were identified, and a simple restriction fragment polymorphism analysis was developed for genotyping. These data provide evidence that GBV-C/HGV consists of three different genotypes. Our simple genotyping assay will also provide a tool for epidemiological studies of GBV-C/HGV infection.

Base Sequence↗