Detection of Single Nucleotide Polymorphisms in p53 Mutation
Hotspots and Expression of Mutant p53 in Human Cell Lines Using
an Enzyme-Linked Electrochemical Assay

J 2009

Detection of Single Nucleotide Polymorphisms in p53 Mutation Hotspots and Expression of Mutant p53 in Human Cell Lines Using an Enzyme-Linked Electrochemical Assay

HORAKOVA, Petra; Eva ŠIMKOVÁ; Zdenka DUDOVÁ; Marie BRÁZDOVÁ; Miroslav FOJTA et al.

Základní údaje

Originální název

Detection of Single Nucleotide Polymorphisms in p53 Mutation Hotspots and Expression of Mutant p53 in Human Cell Lines Using an Enzyme-Linked Electrochemical Assay

Autoři

HORAKOVA, Petra; Eva ŠIMKOVÁ; Zdenka DUDOVÁ; Marie BRÁZDOVÁ a Miroslav FOJTA

Vydání

Electroanalysis, WEINHEIM, WILEY-VCH Verlag GmbH, 2009, 1040-0397

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 2.630

Označené pro přenos do RIV

DOI

https://doi.org/10.1002/elan.200904656

UT WoS

000268790300010

Klíčová slova anglicky

Enzyme-linked electrochemical assay; SNP typing; p53; Mutation; Magnetic beads; Polymorphism

Změněno: 28. 6. 2016 14:45, Mgr. Zdenka Dudová, Ph.D.

Anotace

V originále

An enzyme-linked electrochemical technique for single nucleotide polymorphism (SNP) typing in the p53 tumor suppres or gene is presented. The technique is based on a DNA polymerase-catalyzed extension of a primer hybridized to a target DNA strand upstream (5'-> 3') to the SNP site by one nucleotide bearing a biotin tag. Under optimized conditions, efficient incorporation of the biotinylated nucleotide occurs only in the case of complementarity between the first nucleotide in single-stranded 5'-overhang of the target strand. The introduced biotin tag is detected after capture of the primer extension products at magnetic beads bearing oligoT strands via oligoA adaptors at 5'-ends of the primer, binding of streptavidin-alkaline phosphatase conjugate and enzymatic conversion of I-naphthyl phosphate into I-naphthol which is determined electrochemically at carbon electrodes. In addition to model studies with synthetic oligonucleotides, we report on detection of mutant p53 expression in human cell lines using reverse transcription-PCR technique combined with amplified primer extension and the magnetic beads-based electrochemical assay.

Citovat

HORAKOVA, Petra; Eva ŠIMKOVÁ; Zdenka DUDOVÁ; Marie BRÁZDOVÁ a Miroslav FOJTA. Detection of Single Nucleotide Polymorphisms in p53 Mutation Hotspots and Expression of Mutant p53 in Human Cell Lines Using an Enzyme-Linked Electrochemical Assay. Electroanalysis. WEINHEIM: WILEY-VCH Verlag GmbH, 2009, roč. 21, č. 15, s. 1723-1729. ISSN 1040-0397. Dostupné z: https://doi.org/10.1002/elan.200904656.

@article{1348786,
   author = {Horakova, Petra and Šimková, Eva and Dudová, Zdenka and Brázdová, Marie and Fojta, Miroslav},
   article_location = {WEINHEIM},
   article_number = {15},
   doi = {https://doi.org/10.1002/elan.200904656},
   keywords = {Enzyme-linked electrochemical assay; SNP typing; p53; Mutation; Magnetic beads; Polymorphism},
   language = {eng},
   issn = {1040-0397},
   journal = {Electroanalysis},
   title = {Detection of Single Nucleotide Polymorphisms in p53 Mutation Hotspots and Expression of Mutant p53 in Human Cell Lines Using an Enzyme-Linked Electrochemical Assay},
   volume = {21},
   year = {2009}
}

TY  - JOUR
ID  - 1348786
AU  - Horakova, Petra - Šimková, Eva - Dudová, Zdenka - Brázdová, Marie - Fojta, Miroslav
PY  - 2009
TI  - Detection of Single Nucleotide Polymorphisms in p53 Mutation Hotspots and Expression of Mutant p53 in Human Cell Lines Using an Enzyme-Linked Electrochemical Assay
JF  - Electroanalysis
VL  - 21
IS  - 15
SP  - 1723-1729
EP  - 1723-1729
PB  - WILEY-VCH Verlag GmbH
SN  - 10400397
KW  - Enzyme-linked electrochemical assay
KW  - SNP typing
KW  - p53
KW  - Mutation
KW  - Magnetic beads
KW  - Polymorphism
N2  - An enzyme-linked electrochemical technique for single nucleotide polymorphism (SNP) typing in the p53 tumor suppres or gene is presented. The technique is based on a DNA polymerase-catalyzed extension of a primer hybridized to a target DNA strand upstream (5'-> 3') to the SNP site by one nucleotide bearing a biotin tag. Under optimized conditions, efficient incorporation of the biotinylated nucleotide occurs only in the case of complementarity between the first nucleotide in single-stranded 5'-overhang of the target strand. The introduced biotin tag is detected after capture of the primer extension products at magnetic beads bearing oligoT strands via oligoA adaptors at 5'-ends of the primer, binding of streptavidin-alkaline phosphatase conjugate and enzymatic conversion of I-naphthyl phosphate into I-naphthol which is determined electrochemically at carbon electrodes. In addition to model studies with synthetic oligonucleotides, we report on detection of mutant p53 expression in human cell lines using reverse transcription-PCR technique combined with amplified primer extension and the magnetic beads-based electrochemical assay.
ER  -

HORAKOVA, Petra; Eva ŠIMKOVÁ; Zdenka DUDOVÁ; Marie BRÁZDOVÁ a Miroslav FOJTA. Detection of Single Nucleotide Polymorphisms in p53 Mutation Hotspots and Expression of Mutant p53 in Human Cell Lines Using an Enzyme-Linked Electrochemical Assay. \textit{Electroanalysis}. WEINHEIM: WILEY-VCH Verlag GmbH, 2009, roč.~21, č.~15, s.~1723-1729. ISSN~1040-0397. Dostupné z: https://doi.org/10.1002/elan.200904656.

Přehled o publikaci