56 resultados para Mutation testing


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枯草芽孢杆菌BJ1是一种在真菌病害防治中发挥重要作用的生防因子,为进一步提高它的抑菌能力,获得生防效果更好的高效菌种,利用不同能量和剂量的12C6+对生防菌BJ1进行了离子辐照处理。研究结果表明:离子辐照生防菌BJ1的最适宜剂量为200~400 Gy,传能线密度(LET)为60 keV/μm;突变菌株的抑菌能力比BJ1提高了2%~21%;不仅防病效果比BJ1提高了17.48%,而且对植物具有更好的促生长作用。

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选用12C6+离子辐照诱变阿维菌素B1a产生菌ZJAV-A1,研究其诱变效应。实验结果表明,12C6+离子辐照剂量50Gy时致死率97%,正突变率最高可达到34.2%。通过12C6+离子诱变处理,结合平板培养基及斜面培养基的正突变菌株筛选,最终获得一株稳定性良好,阿维菌素B1a组分产量稳定在4460—4588μg/ml之间,较出发菌株提高11.1%—14.7%的突变株ZJAV-Y1-203。

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Several methods of mutation detection, such as single-strand conformation polymorphism (SSCP), tandem SSCP/heteroduplex analysis and SNaPshot analysis were developed using homemade kit on ABI 310 genetic analyzer, and were successfully applied to mutation detection of 31 colorectal tumor samples. The sieving capability of homemade kit and commercial kit were compared, results demonstrate that homemade kit has higher resolution and shorter analysis time. In clinical tumor samples, 26% K-ras (exon 1) and 24% p53 (exons 7-8) were found to have mutations, and all mutations were single point variations. A majority of mutations occurred in one gene, only 1 tumor contained alterations in the two genes, which indicates that development of colorectal cancer lies on alternate pathways, and may correlate with different gene mutations.

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Mutation of hMLH1 gene plays an important role in human tumorigenesis. A highly sensitive single-strand conformation polymorphism (SSCP) method for detection of the T1151A mutation in exon 12 of the hMLH1 gene was for the first time developed employing laser-induced fluorescence capillary electrophoresis (LIF-CE). Effects of the concentration of linear polyacrylamide solution, running temperature, running voltage and the addition of glycerol on SSCP analysis were investigated, and the optimum separation conditions were defined. Thirty colorectal cancer patients and eight lung cancer patients were screened and the T1151A mutation was found in four of them. Based on CE-sequencing the mutation was further confirmed. To our knowledge, this is for the first time that the T1151A mutation is found in lung cancer. Our method is simple, rapid, and highly sensitive and is well suited to the analysis of large numbers of clinical samples.

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The Charpy impact fracture behaviour of unnotched specimens of phenolphthalein polyether ketone (PEK-C) was studied over a temperature range from room temperature to 220 degrees C by using an instrumented impact tester. The load-time and energy-time curves of PEK-C at different temperatures were recorded. From these curves, some important parameters, such as the maximum impact load, the maximum stress, the total impact energy, the crack initiation energy, the crack propagation energy etc., were obtained and their temperature dependences of PEK-C were investigated. The point of 100 percent maximum load on the load-time trace was shown to be the yield point. Two parameters, the ductile ratio (D.R.) and the ductility index (D.I.) were applied to characterize the ductility of PEK-C and their relationships to the relaxation processes were discussed.

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A continuous spatial temperature gradient was established in capillary electrophoresis by using a simple temperature control device. The temperature profile along the capillary was predicted by theoretical calculations. A nearly linear spatial temperature gradient was established and applied to DNA mutation detection. By spanning a wide temperature range, it was possible to perform simultaneous heteroduplex analysis for various mutation types that have different melting temperatures.

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A supported lanthanum gallate (LSGM) electrolyte thin-film solid oxide fuel cell with Ni-YSZ cermet anode and strontium-doped lanthanum manganite (LSM)-yttria stabilized zirconia (YSZ) composite cathode was, for the first time, fabricated and tested. The cell was prepared by an unconventional approach, in which an LSGM thin film (about 15 mum thick) was first deposited on a porous substrate such as a porous YSZ disk by a wet process and sintered at a high temperature (above 1400degrees C). NiO was then incorporated into the porous substrate by a carefully controlled impregnation process and fired at a much lower temperature. In this way, the severe reaction between LSGM and NiO at a high temperature, which is required for the full densification of LSGM film, can be avoided. A strontium-doped LaMnO3 (LSM)-YSZ composite cathode was screen printed on the surface of the LSGM film and then fired at 1250degrees C. The electrolyte resistances of the SOFC single cells fabricated by this approach are much lower compared to those of thick LSGM film supported cells. A maximum output power density of over 0.85 W/cm(2) at 800degreesC with H-2 as fuel and air as oxidant for a fabricated cell was achieved. (C) 2002 The Electrochemical Society.

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In this study, an in vitro multicellular tumor spheroid model was developed using microencapsulation, and the feasibility of using the microencapsulated. multicellular tumor spheroid (MMTS) to test the effect of chemotherapeutic drugs was investigated. Human MCF-7 breast cancer cells were encapsulated in alginate-poly-L-lysine-alginate (APA) microcapsules, and a single multicellular spheroid 150 mu m in diameter was formed in the microcapsule after 5 days of cultivation. The cell morphology, proliferation, and viability of the MMTS were characterized using phase contrast microscopy, BrdU-Iabeling, MTT stain, calcein AM/ED-2 stain, and H&E stain. It demonstrated that the MMTS was viable and that the proliferating cells were mainly localized to the periphery of the cell spheroid and the apoptotic cells were in the core. The MCF-7 MMTS was treated with mitomycin C (MC) at a concentration of 0.1, 1, or 10 times that of peak plasma concentration (ppc) for up to 72 h. The cytotoxicity was demonstrated. clearly by the reduction in cell spheroid size and the decrease in cell viability. The MMTS was further used to screen the anticancer effect of chemotherapeutic drugs, treated with MC, adriamycin (ADM) and 5-fluorouracil (5-FU) at concentrations of 0.1, 1, and 10 ppc for 24, 48, and 72 h. MCF-7 monolayer culture was used as control. Similar to monolayer culture, the cell viability of MMTS was reduced after treatment with anticancer drugs. However, the inhibition rate of cell viability in MMTS was much lower than that in monolayer culture. The MMTS was more resistant to anticancer drugs than monolayer culture. The inhibition rates of cell viability were 68.1%, 45.1%, and 46.8% in MMTS and 95.1%, 86.8%, and 91.6% in monolayer culture treated with MC, ADM, and 5-FU at 10 ppc for 72 h, respectively. MC showed the strongest cytotoxicity in both MMTS and monolayer, followed by 5-FU and ADM. It demonstrated that the MMTS has the potential to be a rapid and valid in vitro model to screen chemotherapeutic drugs with a feature to mimic in vivo three-dimensional (3-D) cell growth pattern.

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AIM: To study the status of hMLH1 gene point mutations of gastric cancer kindreds and gastric cancer patients from northern China, and to find out gene mutation status in the population susceptible to gastric cancer.