Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.7 (DNA polymerase)
 17,007 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Enzyme inhibition by fullerene derivatives has attracted much attention. In this communication, effects of two water-solube fullerene derivatives, fullerol and trimalonic acid C60 (TMA C60) on polymerase chain reaction (PCR) were investigated by using PCR of beta-actin cDNA derived from HeLa cells as an experimental model. Both fullerol and TMA C60 were found to inhibit PCR in a dose-dependent manner. PCR was ultimately inhibited while the concentrations of each compound were not less than 0.01 mM. In contrast, mannitol exerted no effects on PCR while its concentration increased up to 2 mM. Compensation experiments with Thermus aquaticus (Taq) DNA polymerase revealed that both fullerol and TMA C60 inhibited the enzymatic activity of Taq DNA polymerase, and the inhibitory potency of TMA C60 was slightly greater than that of fullerol. Our data provides some novel aspects on the enzyme inhibiting activities of fullerene derivatives.
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PMID:Inhibition of a thermophilic deoxyribonucleic acid polymerase by fullerene derivatives. 1767 10

Novel biological activities of a fullerene-oligonucleotide conjugate (FONC) were examined in the present report. A new FONC containing a complementary sequence to a specific region of the beta-actin cDNA was synthesized. We found that the FONC at a low concentration (10(-6)M) could specifically inhibit PCR-amplification of the beta-actin cDNA by inhibiting the activities of both Taq DNA polymerase and the cDNA template. This inhibition appeared to be mediated by reactive oxygen species (ROS). The FONC also exhibited the ability of antagonizing the enzymatic activity of an engineered nuclease, exonuclease I (Exo I). Our data provided some valuable evidences on the prospective use of FONC in the antisense strategy for gene silencing.
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PMID:Inhibition of in vitro amplification of targeted DNA fragment and activity of exonuclease I by a fullerene-oligonucleotide conjugate. 1845 29

The polymerase chain reaction (PCR) theoretically offers a very powerful means of quantifying gene expression in cells and tissues of different histological classes. From a technical viewpoint however, the use of RT-PCR to quantify gene expression can be demanding with poor reproducibility arising from several diverse sources. In this study, we describe and fully characterise an RT-PCR assay which generates highly reproducible estimates of DNA polymerase beta gene expression relative to the expression of beta-actin in a semi-quantitative manner. Particular emphasis has been placed on the efficiency of first strand cDNA synthesis and to aspects of primer design. In addition, various aspects associated with the quantification of gene expression required to generate reproducible results are discussed. Using the techniques described herein, the quantification of DNA polymerase beta expression in three independent experiments using human colon carcinoma cells was highly reproducible with ratios of target to internal standard gene expression of 3.68x10(-4) +/- 0.23x10(-4). Provided that careful consideration is given to key areas of the RT-PCR assay during experimental work up procedures, this assay can be used to provide an accurate measure of gene expression in cell lines.
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PMID:Semiquantitative measurement of gene-expression by rt-PCR - a cautionary tale. 2157 79

Herpesvirus (HV) infections in cetaceans are frequently associated with skin and mucosal lesions. Although HV infections have been reported worldwide, their occurrence in southern Atlantic marine mammals is still poorly understood. We tested skin, oral and genital mucosal beta-actin PCR-positive samples from 109 free-ranging Brazilian cetaceans using a universal herpesvirus DNA polymerase PCR. Herpesvirus-positive skin samples from a Guiana dolphin (Sotalia guianensis), a dwarf sperm whale (Kogia sima), a Bolivian river dolphin (Inia boliviensis), and a lingual sample from an Atlantic spotted dolphin (Stenella frontalis) were histologically evaluated. Additional tissue samples from these animals were also PCR-positive for HV, including a novel sequence obtained from the dwarf sperm whale's stomach and mesenteric lymph node. Four novel HV species were detected in the Guiana dolphin (one), the dwarf sperm whale (two) and the Bolivian river dolphin (one). The cutaneous lesions (marked, focally extensive, chronic proliferative dermatitis) of the Guiana dolphin and the Bolivian river dolphin were similar to previous HV reports in cetaceans, despite the absence of intranuclear inclusion bodies. This is the largest HV survey in South American cetaceans and the first detection of HV infection in riverine dolphins worldwide.
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PMID:Novel herpesviruses in riverine and marine cetaceans from South America. 3046 43


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