PubMed Health⌕ Search

Biomedical subjects

Maria Sasvari-Szekely

Publications and source records attributed to Maria Sasvari-Szekely.

23 records · Page 2Linked to original sources
PubMed2003

Transcription factor binding study by capillary zone electrophoretic mobility shift assay.

Regulation of gene expression through interaction of proteins with specific DNA sequences is a central issue in functional genomics. Capillary electrophoretic mobility shift assay is an efficient novel method for the investigation of sequence specific protein-DNA interactions, allowing rapid and sensitive quantification of the complex formation. In this paper, we present a pilot study on capillary zone electrophoretic mobility shift assay (CZEMSA) to investigate the interaction between the transcription factors of HeLa nuclear extract and Sp1-specific fluorescein-labeled oligonucleotide, using the unlabeled probe as competitor. The mobility shift assay was accomplished by CZE in coated capillaries without polymeric buffer additives. Specificity of the DNA protein complex formation was verified by competition experiments, as well as by supershift assay with an anti-Sp1 antibody. The applied electric field strength did not affect the stability of DNA-protein complex during the electrophoretic analysis, allowing rapid identification and quantification of the protein DNA interaction. A practical application to study the interaction between Oryza sativa MADS-box transcription factor 4 (OsMADS4) and its consensus sequence is also reported.

Base Sequence↗
PubMed2002

Direct haplotype detection of adjacent polymorphic sites in the regulatory region of the dopamine D4 receptor (DRD4) gene.

A novel method of haplotype identification is discussed allowing simultaneous detection of two adjacent polymorphic sites, a single nucleotide polymorphism (SNP) and a length polymorphism within 1-2 kilobase distance. The method combines allele specific-amplification with high-throughput, automated ultrathin-layer gel electrophoresis analysis of fragment size polymorphism. A typical application is shown for genotyping the -521 C/T single nucleotide polymorphism and the 120 bp duplication in the 5"-upstream region of the dopamine D4 receptor (DRD4) gene. We have also demonstrated that the haplotypes of double heterozygotes for -521 C/T and for the 120 bp duplication can be clearly distinguished, that has only been possible previously by extensive pedigree analysis.

Binding Sites↗
PubMed2002

Membrane-mediated ultrafast restriction digestion and subsequent rapid gel microchip electrophoresis of DNA.

Ultrafast, membrane-mediated restriction digestion of DNA molecules followed by rapid gel microchip electrophoresis of the resulting fragments is described. Combination of restriction endonuclease digestion on small pore-size microfibrous membranes with sample loading and electrophoresis analysis in a multilane (up to 96) format resulted in very fast restriction digest based microscale DNA analysis. Complete digestion of several nanogram target DNA was accomplished on the microporous membrane at room temperature just in a few minutes with a single or a combination of various restriction enzymes, using only submicroliter quantities of samples and reagents. The reaction mixture containing membrane also served as sample loading device for the subsequent gel microchip electrophoresis based analysis. This work establishes methods for high-speed, high-throughput DNA analysis, featuring extremely low sample and reagent consumption, and fast restriction digestion in combination with sample loading and rapid gel microchip analysis of the resulting fragments. The entire restriction digestion, sample loading and electrophoresis analysis process required less than 20 min.

Bacteriophage phi X 174↗
PubMedDate not supplied

High-throughput genotyping by microchip electrophoresis.

Easy applicability of modern microfabrication technology to electrophoresis microchips has initiated a rapidly moving interdisciplinary field in analytical chemistry. Electric field-mediated separations in microfabricated devices are significantly faster than conventional electrophoresis methods and are usually completed in seconds to minutes. The flexibility of fluidic manipulations in electrophoresis microchips allows the use of a variety of separation techniques and conditions. In this study, large-scale genotyping of the repeat polymorphism in the regulatory (promoter) region of the serotonin transporter gene 5-HTT linked polymorphic region (5-HTTLPR) was attempted using polymerase chain reaction (PCR) amplification followed by rapid microchip electrophoresis analysis of the amplicons.

DNA↗