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1.
Biosci Biotechnol Biochem ; 84(4): 670-677, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31842715

RESUMO

Rapid DNA template preparation directly from a single rice (Oryza sativa) grain or rice flour of its equivalent weight was developed for loop-mediated isothermal amplification (LAMP). LAMP efficiency using DNA extract obtained from consecutive addition of alkaline lysis reagent (25 mM NaOH, 0.2 mM EDTA) and neutralizing reagent (40 mM Tris-HCl [pH 5]) was comparable to that using an equivalent amount of purified DNA as template. The stability of the prepared DNA extract was confirmed for up to six-day storage at room temperature. Without using any special laboratory devices, the developed method enabled a rapid, simple, and low-cost DNA template preparation method for reliable LAMP testing to detect rice genes.

2.
Food Chem ; 305: 125426, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31522124

RESUMO

Genetically modified (GM) Atlantic salmon, AquAdvantage (AquAd), was the first GM animal approved officially for human consumption. Many countries monitor the use of this product under their GM regulations, but a pragmatic system for AquAd-specific detection is needed. Here, we developed a real-time polymerase chain reaction method with high sensitivity for detection of AquAd in foods. This method showed high specificity for the AquAd transgene and the detection limit was 12.5-25 targeted DNA copies per test reaction. An inter-laboratory study using the method developed demonstrated reproducibility at >0.1% (w/w) AquAd content.


Assuntos
Alimentos Geneticamente Modificados , Reação em Cadeia da Polimerase em Tempo Real/métodos , Salmo salar/genética , Alimentos Marinhos/análise , Animais , Animais Geneticamente Modificados , Reprodutibilidade dos Testes
3.
Int Med Case Rep J ; 12: 355-361, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31819668

RESUMO

Purpose: Ipilimumab is an immune checkpoint inhibitor that is now widely used for patients with metastatic malignant melanoma. However, this immune checkpoint inhibitor can induce related adverse events in various organs. Here, we report a case of bilateral drug (ipilimumab)-induced serous retinal detachment (SRD), and the characteristic features found during swept-source optical coherence tomography (SS-OCT). Case: A 78-year-old man with metastatic melanoma received 4 cycles of dacarbazine, nimustine, vincristine, and interferon-beta starting in January 2014, followed by 47 cycles of nivolumab starting in September 2014. Treatment with ipilimumab was started in June 2017. After 2 cycles of ipilimumab, the patient noticed impairment of the visual field in both of his eyes (at day 22) and visited our department in August 2017. Best corrected visual acuity at this initial visit was 16/20 in both eyes, while a fundus examination revealed SRD and retinal pigment epithelial detachments in both eyes. When using the horizontal and vertical 12 mm B-scanned images of SS-OCT, we detected SRD accompanied by a widely distributed high reflection of the photoreceptor outer segment, including the area where the SRD appeared. In addition, the image also showed the interdigitation zone was indistinguishable. Fluorescein angiography showed little leakage in the maculae. During his course of follow-up examinations, there was an increase in the SRD at the beginning, after which it then gradually decreased. However, the obscurity of the interdigitation zone remained. Conclusion: Ipilimumab may cause impairment of the outer retinal layer. We suggest that this is a presumed ipilimumab-induced SRD.

4.
Data Brief ; 27: 104695, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31720342

RESUMO

This article is referred to the research article entitled "Development of a novel method for specific detection of genetically modified Atlantic salmon, AquAdvantage, using real-time polymerase chain reaction" by Soga et al. (2020). Applicability of the developed growth hormone 1 (GH1) and 18S ribosomal DNA (18S rDNA) detection methods using real-time polymerase chain reaction (PCR) for detecting Atlantic salmon (Salmo salar) to processed food commodities was examined. DNAs extracted and purified from 24 commodities labelled to include salmon as an ingredient were used as template. Yield and purity of DNAs obtained and Cq values from real-time PCR analyses were provided.

5.
Chem Pharm Bull (Tokyo) ; 67(10): 1042-1045, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31582624

RESUMO

Biaryls are important compounds with widespread applications in many fields. Tetramethylammonium fluoride tetrahydrate was found to promote the biaryl coupling of aryl iodides bearing electron-withdrawing substituents with unactivated arenes. The reaction takes place at temperatures between 100 and 150°C and can be applied to a wide range of aromatic and heteroaromatic rings, affording the products in moderate to high yields. The reaction does not require strong bases or expensive additives that are employed in the existing methods and can be conducted in air and moisture without any precautions.


Assuntos
Hidrocarbonetos Aromáticos/química , Hidrocarbonetos Iodados/química , Compostos de Amônio Quaternário/química , Ar , Estrutura Molecular , Temperatura Ambiente , Água/química
6.
Nat Methods ; 16(4): 323-325, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30923384

RESUMO

Our method for analyzing histone modifications, scChIC-seq (single-cell chromatin immunocleavage sequencing), involves targeting of the micrococcal nuclease (MNase) to a histone mark of choice by tethering to a specific antibody. Cleaved target sites are then selectively PCR amplified. We show that scChIC-seq reliably detects H3K4me3 and H3K27me3 target sites in single human white blood cells. The resulting data are used for clustering of blood cell types.


Assuntos
Cromatina/química , Histonas/química , Nuclease do Micrococo/química , Animais , Anticorpos/química , Imunoprecipitação da Cromatina , Biologia Computacional , DNA/química , Epigenômica , Genoma , Sequenciamento de Nucleotídeos em Larga Escala , Código das Histonas , Humanos , Leucócitos/citologia , Leucócitos/metabolismo , Masculino , Camundongos , Células NIH 3T3 , Nucleossomos , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , Processamento de Proteína Pós-Traducional , Reprodutibilidade dos Testes , Análise de Sequência de DNA , Software
7.
Anal Sci ; 35(1): 79-83, 2019 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-30449837

RESUMO

The conjugation of biomolecules, such as protein, sugar, and DNA, with metal nanoparticles is an important technique for bioassay and biomaterial preparation. In this study, we aim to enzymatically immobilize a functional peptide on gold nanoparticles (AuNPs) using a single-step reaction. We used tyrosinase, a catechol oxidase, to immobilize an enzymatic peptide. We performed immobilization experiments of a fluorescent compound-linked caspase-3 substrate peptide using tyrosinase on chitosan-coated AuNPs. Peptides were effectively immobilized onto the AuNPs depending on the presence of tyrosine within the sequence, which suggests the DOPA-quinone produced from tyrosine, via tyrosinase, is connected to the chitosan amino group. Although fluorescent emission from the immobilized capase-3 substrate was quenched by AuNPs, fluorescence intensity recovery occurred due to the addition of caspase-3. Thus, we were able to easily prepare functional AuNPs that can be used for a caspase-3 activity assay. Our results indicate that the tyrosinase-mediated peptide link to chitosan-coated particles is a useful technique for preparing functionalized nanoparticles.


Assuntos
Quitosana/química , Cloretos/química , Enzimas Imobilizadas/química , Compostos de Ouro/química , Nanopartículas Metálicas/química , Monofenol Mono-Oxigenase/química , Peptídeos/química , Corantes Fluorescentes/química , Medições Luminescentes
8.
J Biochem ; 164(6): 407-414, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30101295

RESUMO

An active recombinant human pancreatic lipase (recHPL) was successfully prepared for the first time from the Escherichia coli expression system using short Strep-tag II (ST II). The recHPL-ST II was solubilized using 8 M urea from E.coli lysate and purified on a Strep-Tactin-Sepharose column. After refolding by stepwise dialyses in the presence of glycerol and Ca2+ for 2 days followed by gel filtration, 1.8-6 mg of active recHPL-ST II was obtained from 1 L of culture. The recHPL was non-glycosylated, but showed almost equal specific activity, pH-dependency and time-dependent stability compared to those of native porcine pancreatic lipase (PPL) at 37°C. However, the recHPL lost its lipolytic activity above 50°C, showing a lower heat-stability than that of native PPL, which retained half its activity at this temperature.


Assuntos
Lipase/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Animais , Dicroísmo Circular , Suplementos Nutricionais/efeitos adversos , Inibidores Enzimáticos/farmacologia , Terapia de Reposição de Enzimas/efeitos adversos , Estabilidade Enzimática , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/metabolismo , Glicosilação , Temperatura Alta/efeitos adversos , Humanos , Corpos de Inclusão/enzimologia , Corpos de Inclusão/metabolismo , Cinética , Lipase/efeitos adversos , Lipase/antagonistas & inibidores , Lipase/química , Lipase/genética , Lipase/isolamento & purificação , Oligopeptídeos/química , Oligopeptídeos/genética , Oligopeptídeos/isolamento & purificação , Oligopeptídeos/metabolismo , Orlistate/farmacologia , Conformação Proteica , Processamento de Proteína Pós-Traducional , Redobramento de Proteína , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Solubilidade , Sus scrofa
9.
Shokuhin Eiseigaku Zasshi ; 59(3): 151-156, 2018.
Artigo em Japonês | MEDLINE | ID: mdl-30033993

RESUMO

Highly processed foods, including soy sauce, cornflakes, starch sugar, beet sugar and vegetable oil, are not currently subject to genetically modified (GM) food labeling, because DNA could not be detected in these food products. Here we re-examined the method of DNA extraction from starch syrup, beet sugar and vegetable oil using commercially available DNA extraction kits. We found that DNA was not stably detected by PCR targeting a species-specific endogenous plant gene. The reason for this may have been that the DNA yield was below the detection limit, because PCR inhibition was not observed.


Assuntos
DNA de Plantas/análise , Análise de Alimentos/métodos , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase
10.
J Agric Food Chem ; 66(29): 7839-7845, 2018 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-29949351

RESUMO

We developed a novel loop-mediated isothermal amplification (LAMP)-based detection method using lateral flow dipstick chromatography for genetically modified (GM) soybean and maize events. The single-stranded tag hybridization (STH) for the chromatography printed-array strip (C-PAS) system was used for detections targeting the cauliflower mosaic virus 35S promoter, mannose-6-phosphate isomerase gene, Pisum sativum ribulose 1, 5-bisphosphate carboxylase terminator, a common sequence between the Cry1Ab and Cry1Ac genes, and a GA21-specific sequence. The STH C-PAS system was applicable for multiplex analyses to perform simultaneous detections. The limit of detection was 0.5% or less for each target. By using the developed method, the LAMP amplification was visually detected. Moreover, the detection could be carried out without any expensive instruments, even for the DNA amplification steps, by virtue of the isothermal reaction. We demonstrated that the rapid and useful method developed here would be applicable for screening GM crops.


Assuntos
Técnicas de Amplificação de Ácido Nucleico/métodos , Plantas Geneticamente Modificadas/genética , Soja/genética , Zea mays/genética , Produtos Agrícolas/química , Produtos Agrícolas/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/química , Soja/química , Zea mays/química , Zea mays/metabolismo
11.
Sci Rep ; 8(1): 8660, 2018 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-29930289

RESUMO

Humans have domesticated many kinds of animals in their history. Dogs and horses have particularly close relationships with humans as cooperative partners. However, fewer scientific studies have been conducted on cognition in horses compared to dogs. Studies have shown that horses cross-modally distinguish human facial expressions and recognize familiar people, which suggests that they also cross-modally distinguish human emotions. In the present study, we used the expectancy violation method to investigate whether horses cross-modally perceive human emotions. Horses were shown a picture of a human facial expression on a screen, and they then heard a human voice from the speaker before the screen. The emotional values of the visual and auditory stimuli were the same in the congruent condition and different in the incongruent condition. Horses looked at the speaker significantly longer in the incongruent condition than in the congruent condition when they heard their caretaker's voices but not when they heard the stranger voice. In addition, they responded significantly more quickly to the voice in the incongruent condition than in the congruent one. To the best of our knowledge, this is the first study to show that horses cross-modally recognized the emotional states of their caretakers and strangers.


Assuntos
Vínculo Homem-Animal de Estimação , Emoções , Cavalos/psicologia , Animais de Estimação/psicologia , Voz , Estimulação Acústica/métodos , Animais , Percepção Auditiva/fisiologia , Feminino , Cavalos/fisiologia , Humanos , Japão , Masculino , Animais de Estimação/fisiologia , Estimulação Luminosa/métodos , Percepção Visual/fisiologia
12.
Yakugaku Zasshi ; 138(1): 39-46, 2018.
Artigo em Japonês | MEDLINE | ID: mdl-29311464

RESUMO

The chemical labeling of proteins with synthetic probes is a key technique used in chemical biology, protein-based therapy, and material science. Much of the chemical labeling of native proteins, however, depends on the labeling of lysine and cysteine residues. While those methods have significantly contributed to native protein labeling, alternative methods that can modify different amino acid residues are still required. Herein we report the development of a novel methodology of tyrosine labeling, inspired by the luminol chemiluminescence reaction. Tyrosine residues are often exposed on a protein's surface and are thus expected to be good targets for protein functionalization. In our studies so far, we have found that 1) hemin oxidatively activates luminol derivatives as a catalyst, 2) N-methyl luminol derivative specifically forms a covalent bond with a tyrosine residue among the 20 kinds of natural amino acid residues, and 3) the efficiency of tyrosine labeling with N-methyl luminol derivative is markedly improved by using horseradish peroxidase (HRP) as a catalyst. We were able to use molecular oxygen as an oxidant under HRP/NADH conditions. By using these methods, the functionalization of purified proteins was carried out. Because N-methyl luminol derivative is an excellent protein labeling reagent that responds to the activation of peroxidase, this new method is expected to open doors to such biological applications as the signal amplification of HRP-conjugated antibodies and the detection of protein association in combination with peroxidase-tag technology.


Assuntos
Tirosina/química , Catálise , Heme/química , Hemeproteínas/química , Luminescência , Luminol/química , Peroxidase/química
13.
Sci Rep ; 7(1): 14942, 2017 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-29097736

RESUMO

Poisonous Entoloma rhodopolium and other similar species including edible E. sarcopum are morphologically diverse. People mistake poisonous species for edible species. Classification and the detection method of these species need to be defined. The morphological and phylogenetic studies have been reported in northern Europe. In Japan, the genetic study remains unsolved. Thus, phylogenetic analysis of E. rhodopolium was conducted using ITS and RPB2 sequences, and the result was compared with that of European species. Japanese E. rhodopolium was classified into three clades, none of which belonged to the true European E. rhodopolium and other known species. Three species were defined as new species. Entoloma rhodopolium clade-I (named E. lacus) was genetically close to but morphologically separated from E. majaloides. Clade-II (E. subrhodopolium) was classified to the same group as E. sinuatum and E. subsinuatum, but distinct from these species. Clade-III was segregated from known Entoloma species including E. lupinum, and named E. pseudorhodopolium. Based on the classification, a simple identification method PCR-RFLP was developed to discriminate between poisonous species and edible E. sarcopum, which is very similar in morphology. The study can help to clarify the taxonomy of complex E. rhodopolium-related species, and to prevent food poisoning.


Assuntos
Agaricales/genética , DNA Fúngico/genética , Polimorfismo de Fragmento de Restrição , Agaricales/classificação , Agaricales/ultraestrutura , Europa (Continente) , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Japão , Filogenia , Especificidade da Espécie
14.
Shokuhin Eiseigaku Zasshi ; 58(3): 113-123, 2017.
Artigo em Japonês | MEDLINE | ID: mdl-28690301

RESUMO

Omphalotus japonicus is a poisonous mushroom that grows in Japan. It can be mistaken for edible mushrooms (Shiitake, Hiratake and Mukitake), and if ingested, it causes food poisoning within 30 min to 1 hr. We established a rapid detection method using PCR-RFLP to identify O. japonicus by restriction digestion of the amplified ITS region. By using Sau96I, Bpu10I, SfcI or DrdI/HincII as a restriction enzyme, it was possible to rapidly identify and discriminate O. japonicus based on the fragment length. This study also provided a short PCR-RFLP system comprising amplification and digestion of a short 200-bp DNA fragment within the ITS region. The system could identify and discriminate O. japonicus after in vitro gastric digestion of native and heated mushroom samples as a model of food poisoning. In addition, a confirmatory assay using real-time PCR was developed to achieve more sensitive detection of O. japonicus.


Assuntos
Agaricales/genética , Agaricales/isolamento & purificação , Análise de Alimentos/métodos , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase em Tempo Real/métodos , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , Humanos , Intoxicação Alimentar por Cogumelos/diagnóstico , Intoxicação Alimentar por Cogumelos/etiologia , Intoxicação Alimentar por Cogumelos/prevenção & controle , Filogenia , Sensibilidade e Especificidade , Fatores de Tempo
15.
Phys Chem Chem Phys ; 19(30): 19707-19721, 2017 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-28530728

RESUMO

Coulomb explosion of diiodomethane CH2I2 molecules irradiated by ultrashort and intense X-ray pulses from SACLA, the Japanese X-ray free electron laser facility, was investigated by multi-ion coincidence measurements and self-consistent charge density-functional-based tight-binding (SCC-DFTB) simulations. The diiodomethane molecule, containing two heavy-atom X-ray absorbing sites, exhibits a rather different charge generation and nuclear motion dynamics compared to iodomethane CH3I with only a single heavy atom, as studied earlier. We focus on charge creation and distribution in CH2I2 in comparison to CH3I. The release of kinetic energy into atomic ion fragments is also studied by comparing SCC-DFTB simulations with the experiment. Compared to earlier simulations, several key enhancements are made, such as the introduction of a bond axis recoil model, where vibrational energy generated during charge creation processes induces only bond stretching or shrinking. We also propose an analytical Coulomb energy partition model to extract the essential mechanism of Coulomb explosion of molecules from the computed and the experimentally measured kinetic energies of fragment atomic ions by partitioning each pair Coulomb interaction energy into two ions of the pair under the constraint of momentum conservation. Effective internuclear distances assigned to individual fragment ions at the critical moment of the Coulomb explosion are then estimated from the average kinetic energies of the ions. We demonstrate, with good agreement between the experiment and the SCC-DFTB simulation, how the more heavily charged iodine fragments and their interplay define the characteristic features of the Coulomb explosion of CH2I2. The present study also confirms earlier findings concerning the magnitude of bond elongation in the ultrashort X-ray pulse duration, showing that structural damage to all but C-H bonds does not develop to a noticeable degree in the pulse length of ∼10 fs.

16.
Chembiochem ; 18(5): 475-478, 2017 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-28009088

RESUMO

The efficiency of protein chemical modification on tyrosine residues with N-methylluminol derivatives was drastically improved by using horseradish peroxidase (HRP). In the previous method, based on the use of hemin and H2 O2 , oxidative side reactions such as cysteine oxidation were problematic for functionalization of proteins selectively on tyrosine residues. Oxidative activation of N-methylluminol derivatives with a minimum amount of H2 O2 prevented the occurrence of oxidative side reactions under HRP-catalyzed conditions. As probes for HRP-catalyzed protein modification, N-methylluminol derivatives showed much higher efficiency than tyramide without inducing oligomerization of probe molecules. Tyrosine modification also proceeded in the presence of ß-nicotinamide adenine dinucleotide (NADH, H2 O2 -free conditions).


Assuntos
Química Click , Peroxidase do Rábano Silvestre/metabolismo , Tirosina/química , Sítios de Ligação , Catálise , Heme/química , Peroxidase do Rábano Silvestre/química , Peróxido de Hidrogênio/química , Modelos Moleculares , Estrutura Molecular
17.
Phys Rev E ; 94(5-1): 052209, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27967080

RESUMO

In the present study, we analyzed field measurement data obtained for a Japanese expressway and used it as a data set for the validation of microscopic simulation models. Consequently, in accordance with previous studies, we confirmed the common features depicted by the fundamental diagram (flux vs density relation) and lane-usage ratio vs density diagram. We found two things regarding lane-changing behavior: (1) a lane change occurs asymmetrically, where a lane change from a slow to a fast lane differs from that from a fast to a slow lane; and (2) the so-called incentive criterion in the case of small gaps between the preceding vehicles in both slow and fast lanes refers to the velocities and /or the relative velocities with respect to the preceding vehicles, whereas that for relatively large gaps refers to the distances to the preceding vehicles is cast into the above incentive criterion in addition to the two factors mentioned above.

18.
Data Brief ; 7: 1165-70, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27408919

RESUMO

This article is referred to research article entitled "Whole genome sequence analysis of unidentified genetically modified papaya for development of a specific detection method" (Nakamura et al., 2016) [1]. Real-time polymerase chain reaction (PCR) detection method for unauthorized genetically modified (GM) papaya (Carica papaya L.) line PRSV-YK (PRSV-YK detection method) was developed using whole genome sequence data (DDBJ Sequenced Read Archive under accession No. PRJDB3976). Interlaboratory validation datasets for PRSV-YK detection method were provided. Data indicating homogeneity of samples prepared for interlaboratory validation were included. Specificity and sensitivity test data for PRSV-YK detection method were also provided.

19.
Cancer Sci ; 107(7): 908-15, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27088640

RESUMO

Asbestos-induced mesothelial carcinogenesis is currently a profound social issue due to its extremely long incubation period and high mortality rate. Therefore, procedures to prevent malignant mesothelioma in people already exposed to asbestos are important. In previous experiments, we established an asbestos-induced rat peritoneal mesothelioma model, which revealed that local iron overload is a major cause of pathogenesis and that the induced genetic alterations are similar to human counterparts. Furthermore, we showed that oral administration of deferasirox modified the histology from sarcomatoid to the more favorable epithelioid subtype. Here, we used i.p. administration of desferal to evaluate its effects on asbestos-induced peritoneal inflammation and iron deposition, as well as oxidative stress. Nitrilotriacetate was used to promote an iron-catalyzed Fenton reaction as a positive control. Desferal significantly decreased peritoneal fibrosis, iron deposition, and nuclear 8-hydroxy-2'-deoxyguanosine levels in mesothelial cells, whereas nitrilotriacetate significantly increased all of them. Desferal was more effective in rat peritoneal mesothelial cells to counteract asbestos-induced cytotoxicity than in murine macrophages (RAW264.7). Furthermore, rat sarcomatoid mesothelioma cells were more dependent on iron for proliferation than rat peritoneal mesothelial cells. Because inflammogenicity of a fiber is proportionally associated with subsequent mesothelial carcinogenesis, iron elimination from the mesothelial environment can confer dual merits for preventing asbestos-induced mesothelial carcinogenesis by suppressing inflammation and mesothelial proliferation simultaneously.


Assuntos
Asbestos/toxicidade , Carcinogênese/efeitos dos fármacos , Desferroxamina/farmacologia , Quelantes de Ferro/farmacologia , Ferro/deficiência , Neoplasias Mesoteliais/induzido quimicamente , Neoplasias Mesoteliais/prevenção & controle , Animais , Peso Corporal , Proliferação de Células/efeitos dos fármacos , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Ferro/química , Ferro/metabolismo , Macrófagos/efeitos dos fármacos , Masculino , Neoplasias Mesoteliais/metabolismo , Neoplasias Mesoteliais/patologia , Ratos , Ratos Wistar
20.
Food Chem ; 205: 272-9, 2016 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-27006240

RESUMO

Identification of transgenic sequences in an unknown genetically modified (GM) papaya (Carica papaya L.) by whole genome sequence analysis was demonstrated. Whole genome sequence data were generated for a GM-positive fresh papaya fruit commodity detected in monitoring using real-time polymerase chain reaction (PCR). The sequences obtained were mapped against an open database for papaya genome sequence. Transgenic construct- and event-specific sequences were identified as a GM papaya developed to resist infection from a Papaya ringspot virus. Based on the transgenic sequences, a specific real-time PCR detection method for GM papaya applicable to various food commodities was developed. Whole genome sequence analysis enabled identifying unknown transgenic construct- and event-specific sequences in GM papaya and development of a reliable method for detecting them in papaya food commodities.


Assuntos
Carica/genética , Frutas/genética , Plantas Geneticamente Modificadas/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Análise de Sequência/métodos , Carica/química , Frutas/química , Genômica
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