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By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus,FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗,本研究通过定点突变方法和PCR扩增方法,获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D,将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接,分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D;对重组质粒进行序列测定、分析,并将重组质粒分别转染BHK-21细胞,通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达,用电子显微镜观察病毒空衣壳的组装;为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果,将重组质粒经肌肉注射方法接种豚鼠,并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫,第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株;随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛,三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus,FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲,本研究通过定点突变方法和PCR扩增方法,获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D,将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接,分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D;对重组质粒进行序列测定、分析,并将重组质粒分别转染BHK-21细胞,通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达,用电子显微镜观察病毒空衣壳的组装;为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果,将重组质粒经肌肉注射方法接种豚鼠,并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫,第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株:随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛,三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

Holding a microscope to the first-mentioned red ant, I saw that,though he was assiduously gnawing at the near fore leg of his enemy,having severed his remaining feeler, his own breast was all torn away, exposing what vitals he had there to the jaws of the black warrior, whose breastplate was apparently too thick for him to pierce; and the dark carbuncles of the sufferer's eyes shone with ferocity such as war only could excite.

用了这显微镜,先来看那最初提起的红蚂蚁,我看到,虽然它猛咬敌人前腿的附近,又咬断了它剩下的触须,它自己的胸部却完全给那个黑色战士撕掉了,露出了内脏,而黑色战士的胸铠却太厚,它没法刺穿;这受难者的黑色眼珠发出了只有战争才能激发出来的凶狠光芒。

MethodsWith nasal endoscopy and microscopy, the middle conchas were moved outside by Hardy′s stretcher, the nasal cavities were broadened, the frontal walls of sphenoid sinus were opened, and the lesions were removed.

方法鼻内镜下用Hardy′s撑开器外移中鼻甲,扩大鼻腔,直达并开放蝶窦前壁,联合显微镜切除病变。

The result seemed fine when viewed with the naked eye, but under the microscope the resolution and contras were poor.

当用肉眼看时,摄影效果似乎不错,但在显微镜下观察时,清晰度和反差都比较差。

The purified vWF was immobilized onto a coverslip. Cell rolling was induced in a parallel-plate flow chamber and observed by phase-contrast video microscope.

纯化的vWF固着在盖玻片上,在平行板液流室中进行细胞滚动研究,用相差电视显微镜观察。

Methods We established the model of human hair follicle in vitro in Williams E serum-free medium. We can continuously observe morphologic change of hair follicle under an invert microscope. The length of hair follicle was measured by eyepiece micrometer and DNA synthesis rate of human hair follicle in vitro by measuring the rates of incorporation of 3H-TdR.Histology form was observed by Cryo-section and transmission electron microscope.

在Williams E无血清培养基中,建立离体人头皮毛囊培养模型;通过倒置显微镜观察毛囊的形态变化;目镜测微器测量毛囊长度;3H-TdR 掺入检测毛囊DNA合成率;组织学检测用冰冻切片光镜观察和电镜切片透射电镜观察。

The morphology and structure of the dendrite-like copper crystals with micrometer size and face-centered cubic structure were characterized by using scanning electron microscopy and X-ray diffraction.

用扫描电子显微镜和X射线衍射仪对枝晶铜进行了形貌和物质结构的表征;制得的枝晶铜尺寸达到微米级,为面心立方晶相。

The AFM study shows the difference in morphology of the polymer fibrils .

用原子力显微镜研究了各样品的聚合物形貌,表明在器件内部形成了沿摩擦方向延伸的聚合物网络,并且随着单体含量的增加聚合物网络更加致密。

The aggregation behaviors of fish scale collagen at different concentration, temperature and pH were studied by AFM.

用原子力显微镜考察了样品质量浓度、溶液pH及温度对鱼鳞胶原蛋白溶液聚集态的影响。

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