查询词典 plasmids

All the probes were pointed on glass slides by GMS 417 Arrayer. Amplified plasmids wer.e labeled by PCR fluorescence(CY5)-primer and CY5-dUTP. The labeled plasmids were applied to the DNA chip. The hybridization was performed at scalar temperatures (43 C, 45 C, 47 C, 49 C, 51C) for one hour. After hybridization the chips were scanned by GMS418 Array Scanner.

采用PCR引物直接标记法和掺入法对HPv标准品进行扩增和荧光标记，将标记好的标准品与芯片在梯度温度下(43℃，45℃，47℃，49℃，51℃)杂交1 小时，用GMs 418 Array Sc~er扫描仪对杂交后的芯片进行扫描检测分析，用 sca'Array软件对各信号点进行信号强度采集。

The plasmids were intradermally injected with a gene gun through 3 bullets, 2μg of plasmids and 4.5Mpa for each shoot.

于质粒注入后24~144h，检测小鼠体内的荧光素酶活性。

Methods:The expressive plasmids of AML1b and AML1-ETO were transfected into CV-1 and 293 cells.The expression level of endogenous pig7 gene was detected by Realtime-PCR.The luciferase reporter plasmids containing pig7 enhancer/corresponding mutant sequences were constructed and co-transfected into CV-1 cells with expressive plasmids of AML1b and AML1-ETO.The transactivity of pig7 enhancer was assayed by luminometer.

研究方法：AML1b和AML7-ETO表达质粒分别转染及共转染CV-1和293细胞，用Realtime-PCR方法检测上述基因对两种细胞内源性pig7基因mRNA表达水平的影响；构建含AML1b结合位点的pig7基因增强子序列以及相对应的突变位点的荧光素酶报告基因质粒；将表达载体/突变载体与AML1b或AML1-ETO基因瞬时共转染CV-1细胞，分析AML1b、AML1-ETO对报告基因的转录调节作用。

After 10 weeks immunization, the combinant plasmids distribute different organs and tissues of the BALB/c mice immunized by DNA vaccines of Toxoplasma gondii ,but only blood was Detected out the recombinant plasmids.

结论弓形虫DNA疫苗免疫BALB／c鼠5周后，多个不同器官组织均可检测出重组质粒，免疫后10周血液检测出重组质粒。

In high dentity fermentation, the cellular respiration, energy metabolism, highest bacterial density and dry bacteria weight of the G830 strain were markedly better than control strains PA1 and BL21, The expression of recombinant prolyl endopeptidase in G830 and PA1 under the above condition was high and stable. Their growth situation and fermentation parameters were similar with their parental strains without plasmids, and resided plasmids maintained stably in those strains.

在高密度发酵条件下，G830的细胞呼吸强度、能量代谢、最高菌密度和细胞干重，均明显优于对照菌株PA1和BL21；重组蛋白脯氨醯内肽酶在G830和PA1中获得稳定高表达；重组菌生长状况及发酵指标均与空宿主菌基本一致且表达质粒能维持较好的稳定性。

However, 2 inserted fragments of recombined plasmids have no overlapped regions. Further more, when these recombined plasmids were retransformted to M145::SCE25AT, no Km~sApr~R and "bald" phenotype transformant were obtained in 5,000 Thio~R transformants.

但是，测定2个重组质粒的插入片段没有发现相互重叠区；而且，提取2株重组质粒重转化到M145：：SCE25AT中，分别得到5，000株Thio~R转化子，但没有得到Km~SApr~R及&光秃&表型的转化子。

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。

Detail Contents: Genetic disorders -- Immune deficiencies -- Breast cancer -- Colon cancer -- Melanoma -- Cystic fibrosis -- Hemophilia -- Liver disease -- Cardiovascular disease -- Muscular dystrophy -- Alzheimer's disease -- Parkinson's disease -- Huntington's disease -- Viruses: the cornerstone of gene therapy -- Viruses are living crystals -- Viral genomes may be RNA or DNA -- Viruses evolved from plasmids -- Viruses know how to infect cells -- The virus as a gene vehicle -- Viruses used in gene therapy -- Ashi DeSilva: a promising start -- Clinical trials defined -- Cells of the immune system -- Adenosine deaminase -- Preliminary research -- Clinical procedure for ADA gene therapy -- The DeSilva clinical trial -- Jesse Gelsinger: down to earth -- Ornithine transcarbamylase -- Preliminary research -- Clinical procedure for OTC gene therapy -- The Gelsinger clinical trial -- The investigation -- Concluding remarks -- Future prospects -- Safer vehicles -- Reducing immune rejection of the vector -- Improved risk assessment -- Redesigning human anatomy and physiology -- Ethics of gene therapy -- The Belmont report -- Clinical trials -- Physiological enhancement -- Cosmetic applications -- Legal issues -- Regulatory agencies -- The Gelsinger legal trial -- International regulation -- Resource center -- Eucaryote cell primer -- Recombinant DNA primer -- The human genome project -- X-linked severe combined immunodeficiency (SCID-X1)-- Alzheimer's disease -- Huntington's disease.

细节内容︰遗传疾病-免疫的缺乏-乳腺癌-结肠癌-黑瘤-囊性纤维变性-血友症-肝疾病-心血管疾病-肌营养不良-早老性痴呆病-帕金森疾病-亨廷顿疾病-病毒︰基础的基因治疗-病毒在活著水晶--病毒的基因可能是RNA或者DNA --病毒从plasmids被逐步形成--病毒知道怎样感染细胞--作为一辆基因车辆的病毒--基因治疗使用的病毒-Ashi DeSilva︰有希望开始-临床试验确定--细胞的这免疫系统-Adenosine deaminase-初步研究-临床程式给埃达基因治疗--这DeSilva临床试验-婕西Gelsinger︰到地球-Ornithine transcarbamylase-初步研究-临床程式给OTC基因治疗-- Gelsinger临床试验-调查-达成评论-前景-更安全的车辆--矢量的降低免疫的拒绝-改进风险估计-重新设计人解剖学和生理学--伦理学的基因治疗-那些贝拉蒙特报告-临床试验-生理提升-美容应用-法律问题-协调机构-- Gelsinger 合法审讯-国际管理-资源中心人物-Eucaryote信元第一-Recombinant DNA 入门--人类基因工程-- X 连结的严重的结合的免疫缺陷(SCID-X1)-早老性痴呆病--亨廷顿的疾病。

Three plasmids named pBSGt pBSGd and pBSGk were constructed respectively with two plasmids named pEGH and pBS305. The plasmid pEGH contained gfp gene and pBS305 is an E. coli -Rhodococcus shuttle vector. These constructed plasmids can be used to detect the efficiency of promoter.

以带有报告基因gfp的质粒pEGH和大肠杆菌-红球菌穿梭载体pBS305为出发质粒，构建了三个带有不同启动子的表达质粒pBSGt、pBSGd和pBSGk，电击转化红平红球菌LY822。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！