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991.
992.
A molecular beacon, bead-based assay for the detection of nucleic acids by flow cytometry 总被引:3,自引:1,他引:2
Horejsh D Martini F Poccia F Ippolito G Di Caro A Capobianchi MR 《Nucleic acids research》2005,33(2):e13
Molecular beacons are dual-labelled probes that are typically used in real-time PCR assays, but have also been conjugated with solid matrices for use in microarrays or biosensors. We have developed a fluid array system using microsphere-conjugated molecular beacons and the flow cytometer for the specific, multiplexed detection of unlabelled nucleic acids in solution. For this array system, molecular beacons were conjugated with microspheres using a biotin-streptavidin linkage. A bridged conjugation method using streptavidin increased the signal-to-noise ratio, allowing for further discrimination of target quantitation. Using beads of different sizes and molecular beacons in two fluorophore colours, synthetic nucleic acid control sequences were specifically detected for three respiratory pathogens, including the SARS coronavirus in proof-of-concept experiments. Considering that routine flow cytometers are able to detect up to four fluorescent channels, this novel assay may allow for the specific multiplex detection of a nucleic acid panel in a single tube. 相似文献
993.
Laura Borello Mario Ferraro Piero Penengo Maria Lia Rossotti 《Biological cybernetics》1981,39(2):79-85
In this paper we propose a model of visual perception in which a positive feedback mechanism can reproduce the pattern stimulus on a neurons screen. The pattern stimulus reproduction is based on informations coming from the spatial derivatives of visual pattern. This information together with the response of the feature extractors provides to the reproduction of the visual pattern as neuron screen electric activity. We simulate several input patterns and prove that the model reproduces the percept. 相似文献
994.
Marcela A. Garabalino Elisa M. Heber Andrea Monti Hughes Sara J. González Ana J. Molinari Emiliano C. C. Pozzi Susana Nievas Maria E. Itoiz Romina F. Aromando David W. Nigg William Bauer Verónica A. Trivillin Amanda E. Schwint 《Radiation and environmental biophysics》2013,52(3):351-361
Boron neutron capture therapy (BNCT) is based on selective accumulation of 10B carriers in tumor followed by neutron irradiation. We previously proved the therapeutic success of BNCT mediated by the boron compounds boronophenylalanine and sodium decahydrodecaborate (GB-10) in the hamster cheek pouch oral cancer model. Based on the clinical relevance of the boron carrier sodium borocaptate (BSH) and the knowledge that the most effective way to optimize BNCT is to improve tumor boron targeting, the specific aim of this study was to perform biodistribution studies of BSH in the hamster cheek pouch oral cancer model and evaluate the feasibility of BNCT mediated by BSH at nuclear reactor RA-3. The general aim of these studies is to contribute to the knowledge of BNCT radiobiology and optimize BNCT for head and neck cancer. Sodium borocaptate (50 mg 10B/kg) was administered to tumor-bearing hamsters. Groups of 3–5 animals were killed humanely at nine time-points, 3–12 h post-administration. Samples of blood, tumor, precancerous pouch tissue, normal pouch tissue and other clinically relevant normal tissues were processed for boron measurement by optic emission spectroscopy. Tumor boron concentration peaked to therapeutically useful boron concentration values of 24–35 ppm. The boron concentration ratio tumor/normal pouch tissue ranged from 1.1 to 1.8. Pharmacokinetic curves showed that the optimum interval between BSH administration and neutron irradiation was 7–11 h. It is concluded that BNCT mediated by BSH at nuclear reactor RA-3 would be feasible. 相似文献
995.
Analysis of Gene Targeting and Intrachromosomal Homologous Recombination Stimulated by Genomic Double-Strand Breaks in Mouse Embryonic Stem Cells 总被引:11,自引:7,他引:11
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To investigate the effects of in vivo genomic DNA double-strand breaks on the efficiency and mechanisms of gene targeting in mouse embryonic stem cells, we have used a series of insertion and replacement vectors carrying two, one, or no genomic sites for the rare-cutting endonuclease I-SceI. These vectors were introduced into the hypoxanthine phosphoribosyltransferase (hprt) gene to produce substrates for gene-targeting (plasmid-to-chromosome) or intrachromosomal (direct repeat) homologous recombination. Recombination at the hprt locus is markedly increased following transfection with an I-SceI expression plasmid and a homologous donor plasmid (if needed). The frequency of gene targeting in clones with an I-SceI site attains a value of 1%, 5,000-fold higher than that in clones with no I-SceI site. The use of silent restriction site polymorphisms indicates that the frequencies with which donor plasmid sequences replace the target chromosomal sequences decrease with distance from the genomic break site. The frequency of intrachromosomal recombination reaches a value of 3.1%, 120-fold higher than background spontaneous recombination. Because palindromic insertions were used as polymorphic markers, a significant number of recombinants exhibit distinct genotypic sectoring among daughter cells from a single clone, suggesting the existence of heteroduplex DNA in the original recombination product. 相似文献
996.
Pla M La Paz JL Peñas G García N Palaudelmàs M Esteve T Messeguer J Melé E 《Transgenic research》2006,15(2):219-228
Maize is one of the main crops worldwide and an increasing number of genetically modified (GM) maize varieties are cultivated
and commercialized in many countries in parallel to conventional crops. Given the labeling rules established e.g. in the European
Union and the necessary coexistence between GM and non-GM crops, it is important to determine the extent of pollen dissemination
from transgenic maize to other cultivars under field conditions. The most widely used methods for quantitative detection of
GMO are based on real-time PCR, which implies the results are expressed in genome percentages (in contrast to seed or grain
percentages). Our objective was to assess the accuracy of real-time PCR based assays to accurately quantify the contents of
transgenic grains in non-GM fields in comparison with the real cross-fertilization rate as determined by phenotypical analysis.
We performed this study in a region where both GM and conventional maize are normally cultivated and used the predominant
transgenic maize Mon810 in combination with a conventional maize variety which displays the characteristic of white grains
(therefore allowing cross-pollination quantification as percentage of yellow grains). Our results indicated an excellent correlation
between real-time PCR results and number of cross-fertilized grains at Mon810 levels of 0.1–10%. In contrast, Mon810 percentage
estimated by weight of grains produced less accurate results. Finally, we present and discuss the pattern of pollen-mediated
gene flow from GM to conventional maize in an example case under field conditions. 相似文献
997.
998.
Summary
Erythroxylum longisetulosum I. Loiola & M. F. Sales and E. timothei I. Loiola & M. F. Sales (sect. Rhabdophyllum) from north-eastern Brazil are described and illustrated, and their taxonomic and ecological relationships are discussed. 相似文献
999.
Lentiviral vectors have been used for gene transfer into the liver but their ability to efficiently transduce quiescent hepatocytes
remains controversial. Lentivirus-mediated gene transfer is more efficient in cycling cells. We determine the effect of H-IL6
in the lentiviral transduction. The lentiviral vector was used to transduce HepG2 cells and mice liver cells, previously treated
with H-IL6. The highest transduction level was observed in HepG2 cells treated with 30 ng/mL H-IL6 and in the mice that received
4 μg H-IL6. Our results suggest that H-IL6 is an inducer of lentiviral gene transfer into the liver cells without any toxicity. 相似文献
1000.
Sergey Ivanov Maria J. Harrison 《The Plant journal : for cell and molecular biology》2014,80(6):1151-1163
Medicago truncatula is widely used for analyses of arbuscular mycorrhizal (AM) symbiosis and nodulation. To complement the genetic and genomic resources that exist for this species, we generated fluorescent protein fusions that label the nucleus, endoplasmic reticulum, Golgi apparatus, trans‐Golgi network, plasma membrane, apoplast, late endosome/multivesicular bodies (MVB), transitory late endosome/ tonoplast, tonoplast, plastids, mitochondria, peroxisomes, autophagosomes, plasmodesmata, actin, microtubules, periarbuscular membrane (PAM) and periarbuscular apoplastic space (PAS) and expressed them from the constitutive AtUBQ10 promoter and the AM symbiosis‐specific MtBCP1 promoter. All marker constructs showed the expected expression patterns and sub‐cellular locations in M. truncatula root cells. As a demonstration of their utility, we used several markers to investigate AM symbiosis where root cells undergo major cellular alterations to accommodate their fungal endosymbiont. We demonstrate that changes in the position and size of the nuclei occur prior to hyphal entry into the cortical cells and do not require DELLA signaling. Changes in the cytoskeleton, tonoplast and plastids also occur in the colonized cells and in contrast to previous studies, we show that stromulated plastids are abundant in cells with developing and mature arbuscules, while lens‐shaped plastids occur in cells with degenerating arbuscules. Arbuscule development and secretion of the PAM creates a periarbuscular apoplastic compartment which has been assumed to be continuous with apoplast of the cell. However, fluorescent markers secreted to the periarbuscular apoplast challenge this assumption. This marker resource will facilitate cell biology studies of AM symbiosis, as well as other aspects of legume biology. 相似文献