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Wyświetlanie 1-15 z 18
Tytuł:
Evaluation of the activity of thermostable DNA polymerases in the presence of heme, as a key inhibitor in the real time PCR method in diagnostics of sepsis
Autorzy:
Gosiewski, Tomasz
Brzychczy-Włoch, Monika
Pietrzyk, Agata
Sroka, Agnieszka
Bulanda, Małgorzata
Powiązania:
https://bibliotekanauki.pl/articles/1039451.pdf
Data publikacji:
2013
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
polymerase inhibitor
heme
real time PCR
sepsis
Opis:
The study aim was evaluation of the usefulness of several thermostable DNA polymerases in real time PCR conducted in the presence of the heme. Our study had the advantage of testing several different polymerases, one of which proved to be the least sensitive to heme activity. We also found that there is no need of supplementing the reaction mixture with protective substances like BSA. Selection of the appropriate polymerase can increase the efficiency of the PCR reaction which is very important for diagnosis of sepsis and for other analyses performed on DNA template isolated from the blood.
Źródło:
Acta Biochimica Polonica; 2013, 60, 4; 603-606
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Potentiometric determination of cysteine with thiol sensitive silver-mercury electrode
Autorzy:
Drożdż, Ryszard
Naskalski, Jerzy
Ząbek-Adamska, Anna
Powiązania:
https://bibliotekanauki.pl/articles/1041141.pdf
Data publikacji:
2007
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
potentiometry
cysteine
real time assay
homocysteine
glutathione
Opis:
A potentiometric procedure for cysteine thiol group concentration monitoring in media generating free radicals was developed using a thiol specific silver-mercury electrode. Electrolytic deposition of mercury on a silver wire and treatment with 20 mM cysteine in 0.5 M NaOH were used to produce the electrode. A silver-chloride electrode in saturated KCl was the reference. A glass capillary with 1 M KNO3 in 1% agarose gel was the liquid junction. The electrode responded to cysteine concentration in the range from 0.01 to 20 mM yielding a perfect linear relationship for the dependence of log [cysteine] versus electrode potential [mV], with b0 (constant) = -373.43 [mV], b1 (slope) = -53.82 and correlation coefficient r2 = 0.97. The electrode potential change per decade of cysteine concentration was 57 mV. The minimal measurable signal response was at a cysteine concentration of 0.01 mM. The signal CV amounted to 4-6% for cysteine concentrations of 0.01 to 0.05 mM and to less than 1% for cysteine concentrations of 0.5 to 20 mM. The response time ranged from about 100 s for cysteine concentrations of 0.01 to 0.1 mM to 30 s at higher cysteine concentrations. The standard curve reproducibility was the best at cysteine concentrations from 0.1 to 20 mM. In a reaction medium containing cysteine and copper(II)-histidine complex ([His-Cu]2+) solution in 55 mM phosphate buffer pH 7.4 the electrode adequately responded to changes in cysteine concentration. Beside cysteine, the silver-mercury electrode responded also to thiol groups of homocysteine and glutathione, however, the Nernst equation slope was about half of that for cysteine.
Źródło:
Acta Biochimica Polonica; 2007, 54, 1; 205-211
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Anticancer activity of new molecular hybrids combining 1,4-naphthalenedione motif with phosphonic acid moiety in hepatocellular carcinoma HepG2 cells
Autorzy:
Długosz, Angelika
Gach, Katarzyna
Szymański, Jacek
Modranka, Jakub
Janecki, Tomasz
Janecka, Anna
Powiązania:
https://bibliotekanauki.pl/articles/1038683.pdf
Data publikacji:
2017
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
MTT test
apoptosis
real-time PCR
flow cytometry
Opis:
Structural motifs found in naturally occurring compounds are frequently used by researchers to develop novel synthetic drug candidates. Some of these new agents are hybrid molecules which are designed through a concept of combining more than one functional element. In this report, anticancer activity of new synthetic molecular hybrids, substituted 3-diethoxyphosphorylnaphtho[2,3-b]furan-4,9-diones and 3-diethoxyphosphorylbenzo[f]indole-4,9-diones, which integrate natural 1,4-naphtalenedione scaffold, present in several anticancer agents, with pharmacophoric phosphonate moiety, were tested against hepatocellular cell line HepG2. Cytotoxicity was examined using MTT assay. Two most potent compounds, furandione 8a and benzoindoldione 12a, which reduced the number of viable HepG2 cells with the IC50 values of 4.13 µM and 5.9 µM, respectively, were selected for further research. These compounds decreased the mRNA expression levels of several genes: Bcl-2, angiogenic vascular endothelial growth factor (VEGF), c-Fos, caspase-8 and increased the expression of Bax, caspase-3 and -9, c-Jun, p21, p53, as determined by quantitative real-time PCR. The ability of these compounds to induce apoptosis and DNA damage was studied by flow cytometry. The obtained data showed that the new compounds inhibited cell viability by increasing apoptosis and decreasing angiogenesis. Compound 8a was a much stronger apoptosis inducer as compared with 12a and strongly activated the intrinsic pathway of apoptosis, associated with the loss of mitochondrial membrane potential and changes in Bax/Bcl-2 ratio. These findings show that the synthetic hybrids combining 1,4-naphthalenedione system and phosphonic acid moiety display potential to be further explored in the development of new anticancer agents.
Źródło:
Acta Biochimica Polonica; 2017, 64, 1; 41-48
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
PCR and real-time PCR assays to detect fungi of Alternaria alternata species
Autorzy:
Kordalewska, Milena
Brillowska-Dąbrowska, Anna
Jagielski, Tomasz
Dworecka-Kaszak, Bożena
Powiązania:
https://bibliotekanauki.pl/articles/1038891.pdf
Data publikacji:
2015
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Alternaria alternata
detection
identification
PCR
real-time PCR
Opis:
Fungi of the Alternaria genus are mostly associated with allergic diseases. However, with a growing number of immunocompromised patients, these fungi, with A. alternata being the most prevalent one, are increasingly recognized as etiological agents of infections (phaeohyphomycoses) in humans. Nowadays, identification of Alternaria spp. requires their pure culture and is solely based on morphological criteria. Clinically, Alternaria infections may be indistinguishable from other fungal diseases. Therefore, a diagnostic result is often delayed or even not achieved at all. In this paper we present easy to perform and interpret PCR and real-time PCR assays enabling detection of A. alternata species. On the basis of alignment of β-tubulin gene sequences, A. alternata-specific primers were designed. DNA from fungal isolates, extracted in a two-step procedure, were used in PCR and real-time PCR assays followed by electrophoresis or melting temperature analysis, respectively. The assays specificity was confirmed, since positive results were obtained for all A. alternata isolates, and no positive results were obtained neither for other molds, dermatophytes, yeast-like fungi, nor human DNA. The assays developed here enable fast and unambiguous identification of A. alternata pathogens.
Źródło:
Acta Biochimica Polonica; 2015, 62, 4; 707-712
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
The correlation analysis of WWOX expression and cancer related genes in neuroblastoma- a real time RT-PCR study
Autorzy:
Nowakowska, Magdalena
Płuciennik, Elżbieta
Wujcicka, Wioletta
Sitkiewicz, Anna
Kazanowska, Bernarda
Zielińska, Elżbieta
Bednarek, Andrzej
Powiązania:
https://bibliotekanauki.pl/articles/1039340.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
neuroblastoma
WWOX gene
LOH
real time RT-PCR
Opis:
Neuroblastoma is one of the most common paediatric cancers, described as unpredictable due to diverse patterns of behaviour. WWOX is a tumour suppressor gene whose expression is reduced in many tumour types. Loss of its expression was shown to correlate with more aggressive disease stage and mortality rate. The aim of this study was to investigate the role of the WWOX tumour suppressor gene in neuroblastoma formation. We performed real-time RT-PCR to analyse levels of WWOX expression in 22 neuroblastic tumour samples in correlation with genes involved in cell cycle regulation (CCNE1, CCND1), proliferation (MKI67), apoptosis (BCL2, BIRC5, BAX) and signal transduction (EGFR, ERBB4). We also evaluated two potential mechanisms - promoter methylation (MethylScreen method) and loss of heterozygosity (LOH) status, which could be connected with regulation of WWOX gene expression. We found a positive correlation between WWOX gene and BCL2 and HER4 JM-a and negative with cyclin D1 and E1. Our observations are consistent with previous findings and emphasise the role of WWOX in cell cycle and apoptosis regulation. Moreover, strong positive association with HER4 JM-a in this tumour type may indicate a role for WWOX in neuroblastoma cell differentiation. The presented results indicate that LOH in locus D16S3096 (located in intron 8) may be involved in the regulation of WWOX mRNAexpression. However, no association between methylation status of WWOX promoter and its expression was observed.
Źródło:
Acta Biochimica Polonica; 2014, 61, 1; 91-97
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Real-time PCR approach in dermatophyte detection and Trichophyton rubrum identification
Autorzy:
Kobylak, Natalia
Bykowska, Barbara
Nowicki, Roman
Brillowska-Dąbrowska, Anna
Powiązania:
https://bibliotekanauki.pl/articles/1039146.pdf
Data publikacji:
2015
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
fungal infections
dermatophytosis
DNA extraction
real-time PCR
Opis:
Dermatophytes are keratinophilic molds that infect human hair, nails and skin. Diagnosis of dermatophytosis is based on morphological, serological and biochemical features. However, identification is difficult and laborious due to similarities between microorganisms. Thus, there is considerable interest to develop mycological diagnostic procedures based on molecular biology methods. In this study, fast, two-step DNA extraction method and real-time PCR was used for detection of dermatophytes DNA using pan-dermatophyte primers and identification of Trichophyton rubrum from pure cultures. The applied method allowed correct detection of all dermatophytes and correct identification of Trichophyton rubrum in less than 2 hours.
Źródło:
Acta Biochimica Polonica; 2015, 62, 1; 119-122
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Usefulness of real-time PCR in long-term follow-up of follicular lymphoma patients
Autorzy:
Tysarowski, Andrzej
Fabisiewicz, Anna
Paszkiewicz-Kozik, Ewa
Kulik, Jadwiga
Walewski, Jan
Siedlecki, Janusz
Powiązania:
https://bibliotekanauki.pl/articles/1041125.pdf
Data publikacji:
2007
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
real-time PCR
follicular lymphoma
minimal residual disease
molecular remission
Opis:
The aim of this study was to evaluate the usefulness of quantitative real-time PCR (RQ-PCR) for the monitoring of molecular remission in follicular lymphoma (FL) patients during long-term follow-up. RQ-PCR by the use of TaqMan® detection system is a sensitive tool to monitor minimal residual disease (MRD) in FL through amplification of the t(14;18) fusion gene during and post-therapy. In most cases the breakpoint region occurs within the major breakpoint region (MBR). Among 75 patients diagnosed with FL, cells harboring the fusion gene BCL2/JH were found in peripheral blood of 31 patients (41%). We further monitored 30 of these patients in a period varying from 6 months to 5 years by RQ-PCR. In our study the level indicating the possibility of the presence of MRD was established at more than five t(14;18)-positive cells in the background of 83000 normal cells. The results of this work also confirmed that the presence of MRD detected by RQ-PCR is an indication for careful observation of patients because of a higher risk of disease recurrence.
Źródło:
Acta Biochimica Polonica; 2007, 54, 1; 135-142
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Cloning and partial characterization of a gene in Bombyx mori homologous to a human adiponectin receptor
Autorzy:
Zhu, Minfeng
Chen, Keping
Wang, Yong
Guo, Zhongjian
Yin, Huijuan
Yao, Qin
Chen, Huiqin
Powiązania:
https://bibliotekanauki.pl/articles/1040735.pdf
Data publikacji:
2008
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
BmAdipoR
bioinformatics study
subcellular localization
real-time quantitative PCR
BmNPV
Opis:
In this study, we report the cloning and characteristics of an adiponectin-like receptor gene from Bombyx mori (BmAdipoR) with highly conserved deduced amino-acid sequences and similar structure to the human adiponectin receptor (AdipoR). Structural analysis of the translated cDNA suggested it encoded a membrane protein with seven transmembrane domains. BmAdipoR was found to be expressed in multiple tissues and highly expressed in Malpighian tubules, fat body and testis. BmNPV (Bombyx mori nucleopolyhedrovirus) bacmid system combined with confocal microscopy revealed that BmAdipoR was targeted to the cell membrane. We also found that infection with BmNPV did not have an effect on BmAdipoR mRNA quantity in the midgut of susceptible Bombyx mori strain (306) at 48 h, but BmAdipoR mRNA quantity increased significantly at 72 h. We concluded that BmAdipoR gene was a membrane protein ubiquitously expressed in Bombyx mori tissues and that its expression was altered by treating with BmNPV.
Źródło:
Acta Biochimica Polonica; 2008, 55, 2; 241-249
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Quantification of thioredoxin mRNA expression in the rat hippocampus by real-time PCR following oxidative stress.
Autorzy:
Yalcin, Ayfer
Powiązania:
https://bibliotekanauki.pl/articles/1041524.pdf
Data publikacji:
2004
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
kainic acid
real-time polymerase chain reaction
oxidative stress
thioredoxin
Opis:
Thioredoxin (Trx) is a multifunctional protein with a redox-active disulfide/dithiol in the active site. Thioredoxin, with its redox-regulating and reactive oxygen species (ROS) scavenging activities, plays several important biologic roles both in intracellular and extracellular compartments. The purpose of this report was to quantify the relative expression of Trx in rat hippocampus following an oxidative stress-involving treatment such as kainic acid (KA) using real-time PCR and the 2-ΔΔCT method. The relative changes in expression of Trx mRNA in KA-treated and control animals were significantly different as 2.02 ± 0.77 and 1.0 ± 0.26, respectively ( P <0.05). Minimum and maximum n-fold changes in Trx expression in KA-treated and control animals were determined as (1.4-5.2) and (0.8-1.3), respectively. Thus, real-time PCR and the 2-ΔΔCT method for data analysis from real-time PCR were found to be an accurate and sensitive method for quantifying Trx mRNA levels.
Źródło:
Acta Biochimica Polonica; 2004, 51, 4; 1059-1065
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Quantification of thioredoxin mRNA expression in the rat hippocampus by real-time PCR following oxidative stress.
Autorzy:
Yalcin, Ayfer
Powiązania:
https://bibliotekanauki.pl/articles/1041529.pdf
Data publikacji:
2004
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
kainic acid
real-time polymerase chain reaction
oxidative stress
thioredoxin
Opis:
Thioredoxin (Trx) is a multifunctional protein with a redox-active disulfide/dithiol in the active site. Thioredoxin, with its redox-regulating and reactive oxygen species (ROS) scavenging activities, plays several important biologic roles both in intracellular and extracellular compartments. The purpose of this report was to quantify the relative expression of Trx in rat hippocampus following an oxidative stress-involving treatment such as kainic acid (KA) using real-time PCR and the 2-ΔΔCT method. The relative changes in expression of Trx mRNA in KA-treated and control animals were significantly different as 2.02 ± 0.77 and 1.0 ± 0.26, respectively ( P <0.05). Minimum and maximum n-fold changes in Trx expression in KA-treated and control animals were determined as (1.4-5.2) and (0.8-1.3), respectively. Thus, real-time PCR and the 2-ΔΔCT method for data analysis from real-time PCR were found to be an accurate and sensitive method for quantifying Trx mRNA levels.
Źródło:
Acta Biochimica Polonica; 2004, 51, 4; 1087-1090
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Physical state of human papillomavirus type 16 in cervical intraepithelial lesions and cancers determined by two different quantitative real-time PCR methods
Autorzy:
Szostek, Slawa
Biesaga, Beata
Zawilinska, Barbara
Klimek, Malgorzata
Kosz-Vnenchak, Magdalena
Powiązania:
https://bibliotekanauki.pl/articles/1038944.pdf
Data publikacji:
2015
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
real-time PCR
human papillomavirus
squamous intraepithelial lesions
cervical carcinoma
Opis:
The aim of this study was to analyse the correlation between a new multiplex qPCR assay and a reference qPCR assay for assessment of the human papillomavirus (HPV16) load and the viral genome status. The study was performed on 100 HPV16 positive samples containing premalignant lesions and carcinomas. HPV16 E2 and E6 gene loads were assessed by two PCR methods. The load of E2 and E6 was normalized to the cell number by qPCR targeting the RNase P open reading frame. The physical state of the viral genome was determined as a ratio of E2/E6 copies number per cell. Among 100 samples analysed, there were no statistically significant differences in the E2 and E6 viral load evaluated by multiplex qPCR and qPCR, the correlation coefficients were 0.98 and 0.97, respectively. There were 19% of samples with the integrated, 73% with mixed and 8% with episomal state of viral genome detected by multiplex qPCR and 17%, 79%, 4%, respectively, found by qPCR. Prevalence of integrated and episomal forms estimated by multiplex qPCR was higher than the one obtained by qPCR (Chi2, p < 0.0001), but in samples with premalignant and malignant diagnoses no significant differences were demonstrated regardless of the methods used. Sensitivity and specificity of multiplex qPCR were 93.7% and 100% as compared with qPCR, the positive predictive value was 100%. In summary, the multiplex qPCR assay in respect of HPV16 load and the frequency of viral genome status was shown to be a sensitive and specific reference method. Simultaneous estimation of E2 and E6 genes in one reaction tube reduces the cost of testing.
Źródło:
Acta Biochimica Polonica; 2015, 62, 4; 923-928
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Detection and differentiation of Newcastle disease virus and influenza virus by using duplex real-time PCR
Autorzy:
Nidzworski, Dawid
Wasilewska, Edyta
Smietanka, Krzysztof
Szewczyk, Bogusław
Minta, Zenon
Powiązania:
https://bibliotekanauki.pl/articles/1039554.pdf
Data publikacji:
2013
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Newcastle disease virus
influenza virus
detection
differentiation
real-time PCR
duplex
Opis:
Newcastle disease virus (NDV), member of the Paramyxoviridae family and avian influenza virus (AIV), member of the Orthomyxoviridae family, are two main avian pathogens causing serious economic problems in poultry health. Both are enveloped, single-stranded, negative-sense RNA viruses and cause similar symptoms, ranging from sub-clinical infections to severe diseases, including decrease in egg production, acute respiratory syndrome, and high mortality. Similar symptoms hinder the differentiation of infection with the two viruses by standard veterinary procedures like clinical examination or necropsy. To overcome this problem, we have developed a new duplex real-time PCR assay for the detection and differentiation of these two viruses. Eighteen NDV strains, fourteen AIV strains, and twelve other (negative control) strains viruses were isolated from allantoic fluids of specific pathogen-free (SPF), embryonated eggs. Four-weeks-old SPF chickens were co-infected with both viruses (NDV - LaSota and AIV - H7N1). Swabs from cloaca and trachea were collected and examined. The results obtained in this study show that by using duplex real-time PCR, it was possible to detect and distinguish both viruses within less than three hours and with high sensitivity, even in case a bird was co-infected. Additionally, the results show the applicability of the real-time PCR assay in laboratory practice for the identification and differentiation of Newcastle disease and influenza A viruses in birds.
Źródło:
Acta Biochimica Polonica; 2013, 60, 3; 475-480
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
The profile of ErbB/Her family genes copy number assessed by real-time PCR in parathyroid adenoma and hyperplasia associated with sporadic primary hyperparathyroidism
Autorzy:
Bednarz, Natalia
Błaut, Krzysztof
Sworczak, Krzysztof
Osęka, Tomasz
Bielawski, Krzysztof
Powiązania:
https://bibliotekanauki.pl/articles/1040637.pdf
Data publikacji:
2009
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
parathyroid hyperplasia
hyperparathyroidism
parathyroid adenoma
ErbB/Her
quantitative real-time PCR
Opis:
Hyperparathyroidism (pHPT) is a relatively frequent endocrinopathy, however, the molecular mechanisms of its etiology remain poorly understood. This disorder is mainly associated with benign tumours (adenoma) and hyperplasia of the parathyroid, hence, the focus is directed also to genes that are likely to be involved in carcinogenesis. Among such genes are ErbB/Her family genes already used in diagnosis of other tumours (e.g., breast carcinoma) and reported also to play a role in development of endocrine lesions. So far, ErbB-1/Her-1/EGFR expression has been detected in pHPT-associated adenomas and hyperplasia as opposed to no expression in normal parathyroid tissue. Moreover, losses or gains of the fragments of chromosomes where ErbB/Her genes are located have been reported. In this study, the gene dosage of ErbB/Her family genes were determined for the first time in parathyroid adenomas, hyperplasia and morphologically unchanged tissue in order to establish their putative role in the development of the disease. Genomic DNA was isolated from 33 patients with sporadic hyperparathyroidism and the gene copy numbers were assessed using real-time PCR. The ErbB/Her genes' profile was unaltered in most of the examined samples. Two low-level amplifications of ErbB-1/Her-1/EGFR gene, two deletions of ErbB-2/Her-2, and six deletions of ErbB-4/Her-4 were found. The ErbB-3/Her-3 gene remained unaffected. No correlation with clinical parameters was found for any gene. Both the low number of alterations and a lack of their associations with clinical parameters exclude the prognostic value of the ErbB/Her genes family in parathyroid tumourigenesis. Nevertheless, the ErbB-4/Her-4 deletions seem to be interesting for further investigations, especially in the context of PTH secretion.
Źródło:
Acta Biochimica Polonica; 2009, 56, 1; 83-88
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Optimization of the Y831C mutation detection in human DNA polymerase gamma by allelic discrimination assay
Autorzy:
Stopińska, Katarzyna
Grzybowski, Tomasz
Malyarchuk, Boris
Derenko, Miroslava
Miścicka-Śliwka, Danuta
Powiązania:
https://bibliotekanauki.pl/articles/1041222.pdf
Data publikacji:
2006
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
real-time PCR
polymerase γ
progressive external ophthalmoplegia
single nucleotide polymorphisms (SNPs)
Opis:
Many well-defined mutations in the gene for the catalytic subunit of polymerase γ (POLG1) have been found to be associated with disease, whereas the status of several mutations remains unresolved due to the conflicting reports on their frequencies in populations of healthy individuals. Here, we have developed a highly sensitive, real-time allelic discrimination assay enabling detection of the Y831C mutation in the POLG1 gene. The Y831C mutation is present in the Polish population at a frequency of 2.25%. The new assay is well suited to both extensive population studies and molecular diagnostics of POLG1.
Źródło:
Acta Biochimica Polonica; 2006, 53, 3; 591-595
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Changes in accumulation of heteroplasmic mitochondrial DNA and frequency of recombination via short repeats during plant lifetime in Phaseolus vulgaris
Autorzy:
Woloszynska, Magdalena
Gola, Edyta
Piechota, Janusz
Powiązania:
https://bibliotekanauki.pl/articles/1039695.pdf
Data publikacji:
2012
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Phaseolus vulgaris
sublimons
heteroplasmy
quantitative real-time PCR
plant mitochondrial genome
mtDNA recombination
Opis:
Recombination via short repeats in plant mitochondrial genomes results in sublimons - DNA molecules with a copy number much lower compared to the main mitochondrial genome. Coexistence of stoichiometrically different mitotypes, called heteroplasmy, plays an important evolutionary role, since sublimons occasionally replace the main genome resulting in a new plant phenotype. It is not clear, how frequency of recombination and sublimon production is regulated and how it is related to changes in the quantity of the main genome and sublimons. We analyzed the accumulation of two recombining main genome sequences and two resulting sublimons in apical meristems, undifferentiated tissues and leaves of different age of Phaseolus vulgaris. Copy numbers of the main genome sequences varied greatly depending on tissue type and organ age while accumulation of sublimons remained much more stable. Although the overall accumulation of plant mtDNA decreased with the leaf age, the quantity of sublimons increased relative to the main genome indicating a higher frequency of recombination via the short 314 bp repeat. Recombination was symmetrical in young developing leaves while in senescent tissues it shifted towards asymmetric events resulting in overrepresentation of one product. We propose that during plant lifetime replication and recombination frequencies change oppositely sustaining heteroplasmic compositions of the genome, which are favorable for inheritance and maintenance of complex plant mtDNA.
Źródło:
Acta Biochimica Polonica; 2012, 59, 4; 703-709
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
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