查询词典 structural gene

Glioma is still one of refractory disease in the neurosurgical field; the development of new primary and adjuvant treatment is vital. Recently, the gene therapy of glioma is developed rapidly and there are many methods about the gene therapy that include: suicide gene therapy, immunologic gene therapy, drug resistangce gene therapy, angiostatin gene therapy and so on. The sucide gene therapy is the most potential approach of antitumer, these nonmammalian genes encode enzyme that convert nontoxic prodrugs into highly toxic metablites. Cells transfected with suicide genes are targeted for specific negative selection, witch can be induced by administrtion of the corresponding produg. Among the enzyme/produg combinations, two of the best characterized system are herpes simplex virus thymidine kinase /ganciclovir and Escherichia coli cytosine deaminase /5-flourocytosine (5-FC). The formor can convert the antiviral nucleoside analogs acyclovir , ganciclovir to their nucleoside monophosphate derivatives, the monophosphate forms are subsequently phosphorylated by endogenus cellular kinases to triphosphates, these molecules are potent inhibitors of DNA synthesis.

近年来脑胶质瘤的基因治疗发展迅速，应运而生的方法有自杀基因、免疫基因、多药耐药基因以及抗血管生成基因等，其中自杀基因被认为是最有前景的基因治疗方法，它又称病毒介导的酶／药物前体疗法，是利用转基因技术将哺乳动物细胞中所不含有的自杀基因转入到哺乳动物肿瘤细胞中，该基因表达的产物可将无毒的药物前体转化为毒性药物，从而选择性杀伤该肿瘤细胞，常用的自杀基因有单纯疱疹病毒-胸苷激酶基因和大肠杆菌胞嘧啶脱氨酶基因，前者催化无毒性抗病毒核苷类似物如丙氧鸟苷、无环鸟苷等成为单磷酸核苷衍生物，然后在内源性细胞激酶作用下转化为具有明显毒性的三磷酸核苷，作为DNA合成链的终止剂和DNA合成酶的抑制剂，干扰细胞DNA的合成；后者编码的胞嘧啶脱氨酶可催化5-氟胞嘧啶（5-FC）脱氨成为5-氟尿嘧啶（5-FU），然后代谢为有毒性的5-氟尿嘧啶-5′三磷酸（5-FUTP）和5-氟-2′脱氧尿嘧啶-5′磷酸（5-FdUTP），5-FUTP通过与UTP竞争性结合而抑制mRNA和tRNA的合成，5-FdUTP则作用于胸苷合成酶，导致TMP衰竭而阻止DNA的合成，最终诱导肿瘤细胞凋亡。

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。

The inhibin a gene、 the bone morphogenetic protein 4 (BMP4) gene and the bone morphogenetic protein 7 (BMP7) gene were studied as candidate genes on the prolificacy of Small Tail Han sheep. Single nucleotide polymorphisms of 5 ' regulatory region、 exon 1 of INHA gene;exon 2、 exon 3 and exon 4 of BMP4 gene and exon 2、 exon 3 of BMP7 were detected in high fecundity breed and low fecundity breeds (Chinese Merino sheep, Corriedale sheep and South African Mutton Merino sheep) by PCR-SSCP. The results indicated that there was a mutation (316C→T) in 5' region of INHA gene in Small Tail Han sheep, Chinese Merino sheep and Corriedale sheep, the same mutation did not exist in South African Mutton Merino sheep.

采用PCR-SSCP技术检测抑制素α(inhibin α，INHA)基因5'调控区、外显子1，骨形态发生蛋白4(bone morphogenetic protein 4，BMP4)基因外显子2、外显子3和外显子4以及骨形态发生蛋白7(bone morphogenetic protein 7，BMP7)基因外显子2和外显子3在高繁殖力绵羊品种以及低繁殖力绵羊品种(中国美利奴绵羊、考力代绵羊、南非肉用美利奴绵羊)中的单核苷酸多态性，同时研究这三个基因对小尾寒羊高繁殖力的影响。

In this experiment, we screen the major protective antigen gene-SOD gene of M. paratuberculosisin order to study the sensitive, specific diagnostic reagent and prophylaxis preparation, especially theDNA vaccine. The SOD gene was amplified from Mycobacterium paratuberculosis C-2 chromosomalDNA by using the PCR technique and cloned into pMD18-T Vector System. We gained a SOD gene of624bp.The recombinant clone was identified byα-complementarity, enzyme digestion and PCRidentification. The result indicated that the recombinant plasmid pMD18-T-SOD was successfullyconstructed. Moreover, through sequential determination and DNASTAR analysis between the clonedSOD gene of M. paratuberculosis C-2 and that of the M.paratuberculosis K-10 strain, the sequentialhomogeneity reached 99%, and the amino acid homogeneity reached 99.5%. The preceding analysisindicated that the SOD gene was very conservative in M. paratuberculosis.

为了研制副结核病敏感、特异的诊断试剂和新型、高效的预防制剂，尤其是DNA疫苗，本研究筛选了M.paratuberculosis主要保护性抗原SOD基因，以M.paratuberculosis C-2染色体DNA为模板，以SOD基因的特异性引物进行PCR扩增，获得了624bp的SOD基因，通过T-A克隆技术，将PCR产物克隆至pMD18-T Vector中，以质粒大小、酶切分析、PCR扩增及序列分析鉴定重组克隆，成功地构建出克隆质粒pMD18-T-SOD，序列测定及DNASTAR分析表明，所获得的M.paratuberculosis C-2 SOD基因与Gen Bank中M.paratuberculosis K-10 SOD基因的大小完全一致，两者核苷酸序列的同源性为99％，氨基酸序列的同源性为99.5％，表明该基因在副结核分枝杆菌中是高度保守的。

After treatment of benzamide which is an inhibitor of poly polymerase,β-galactosidase activity assays and PCR analysis were performed to test the deletion of LacZ gene and c-Ha-T24ras gene. The results showed that:(1)LacZ gene was deleted from both A13.4-NIH3T3 cells and B12.7-NIH3T3 cells, but neigher from A13.4-HeLa cells nor B12.7-HeLa cells. So it is possible that there is cell line specificity in gene deletion induced by BA.(2) In A13.4-NIH3T3 cells, LacZ gene and c-Ha-T24ras gene were completely deleted at the same time.(3) The transcription activity of exgenous DNA fragment may have same effect on its sensitivity to the deletion induced by BA.

用聚ADP核糖聚合酶的NAD位点抑制剂苯甲酰胺分别处理四种转化细胞后，检测细胞中整合的外源LacZ基因与c-Ha-T24ras基因的删除情况，结果如下：BA诱导的基因删除可发生于NIH3T3转化细胞系，但不能发生于HeLa转化细胞系；位于同一外源表达载体上的LacZ基因与c-Ha-T24ras基因的删除过程是同步的；外源整合DNA片段的转录强度可能直接影响其被BA诱导删除的敏感性。

Drug sensitive test and three-dimensional test220 strains of Pa were isolated from hospitalized patients between 2003 and 2007. K-B method was used to tested the susceptibility of 10 different antibiotics. IRPa was screened by testing the minimal inhibitory concentration of imipemem by using agar diluiion method.The susceptibility of these IRPa to the antibiotics was analysised. Three-dimensional test was used to identify the different kinds of beta lactamases from 220 strains of Pa.2.Carbarpenems hydrolytic enzyme genes and oprD2 gene were detectedamong the selected IRPa strains, PCR method was performed to detect carbapenemase genes which included GES、KPC、SPM、VIM、IMP、GIM gene and the oprD2 gene;Multiplex PCR were used to detect OXA genes and plasmid-mediated AmpC beta lactamase genes; The expression of the chromosomal AmpC beta lactamases and oprD2 genes in IRPa strains were analyzed by Real-time PCR.3.Identification and characterization of integronsIntegrase gene was detected by PCR, and the classification of integrons was performed by using restriction fragment length polymorphism.PCR was performed to detect the qacE△1-sull gene,and the gene cassetes which are located at variable region of integrons in the strains were detected to be positive.

方法1、药敏实验和三维实验收集2003～2007年临床分离的220株Pa，对这些菌株采用K-B法测定10种临床常用抗生素的药敏情况，同时采用琼脂稀释法检测亚胺培南的最低抑菌浓度(Minimal inhibitory concentration，MIC)，筛选出对亚胺培南耐药的铜绿假单胞菌，并分析其对其它抗生素的药物敏感率；采用三维实验的方法分析220株Pa产β内酰胺酶的类型。2、碳青霉烯类水解酶和oprD2蛋白的检测针对鉴定的IRPa菌株，采用普通PCR方法检测具有碳青霉烯水解作用的β内酰胺酶耐药基因(GES、KPC、SPM、VIM、IMP、GIM基因)和oprD2基因，采用多重PCR的方法检测OXA型基因和质粒携带的AmpC酶基因，用荧光定量RT-PCR方法检测oprD2蛋白基因表达情况；同时对产AmpC酶的Pa(25株，含IMP耐药和敏感株)用RT-PCR方法检测AmpC酶基因的表达量情况。3。

The experiment obtains clone pig HUMMLC2B gene (GenBank logs onto date: DQ533994), use PCR-RFLP technology, analysed HUMMLC2B gene the 1st embedded child medium Msp Ⅰ is enzymatic cut much condition (T613C) is in distributinging; analysed the much condition sex in 7 breed pig 5 many condition sex and 36 long white pigs, groups 104 pigs and grow to be mixed in vain 5, the result makes clear, the scale that waits for a gene and B to wait for a gene frequency except the A in long white pig in detected swinery is 1 ∶ 2 outside, of the others all take absolutely advantage for a gene such as B, and a gene such as A is pure close individual detect in long white pig only.

实验获得克隆猪HUMMLC2B基因(GenBank登录号：DQ533994)，并采用PCR-RFLP技术，分析了HUMMLC2B基因第1内含子中的MspⅠ酶切多态(T613C)在7个品种猪中的多态性分布；分析了多态性和36头长白猪、5个群体104头猪以及长白和5个群体之和的140头猪胴体性状和肉质性状间的相关，结果表明，在检测的猪群中除长白猪中A等位基因和B等位基因频率的比例为1∶2外，其余的均为B等位基因占绝对优势，且A等位基因纯合个体只在长白猪中检测到。

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。

Based on investigating the short and intermittent structural plane, surrounding rockmass of the huge underground powerhouse in Three Gorge has been divided into five statistical similar areas according to its characteristic of rockmass structure by applying structure plane network simulation. Distributed characteristic of structural plane has been studied and based on it mechanics macrocharacteristic has been studied:(1) It is the first time that based on the result of structural plane network simulation rockmass quality of every structure area has been studied with applying blurred information optimum seeking technique and be compared with and RQD method. The result indicated that it has better effect to evaluate rock quality with blurred information optimum seeking technique.(2) It is the first time that the interrelated relationship of the result of in-suit rockmass deformation test and fractal characteristic of structure on test point has been applied to determine rockmass deformation parameter. The question of evaluating deformation parameters of engineering rockmass has been solved.(3) Anisotropy strength parameters have been determined by applying shear zones simulating method. The result indicated that there are different strength parameters in every structural area, after comparing the result of Fast Lagranian Analysis in continua number simulation method with that of method of weighted mean according to continuity. The result indicated that when a suitable reduction coefficient is been considered rockmass strength parameters can be simply and fast determined with method of weighted mean according to continuity where there is not enough test result.(4) Damage characteristic has been studied based on analysis fractal characteristic of structural plane network by applying fractal-damage coupling method. It is the first time that the concept of damage degree has been set up to evaluate rockmass quality according to the biggest principal damage.

通过对短小、断续性结构面统计特征的研究，将地下厂房区划分为五个岩体结构统计均质区，应用结构面网络模拟技术研究了各个岩体结构均质区结构面的发育分布特征，并以此为基础对岩体宏观力学特性进行了深入研究：(1)首次通过在结构面网络模拟计算结果的基础上，应用模糊信息优化技术研究了厂房区各个岩体结构分区的岩体质量并与岩体基本质量指标研究法、岩体质量指标RQD值研究法作了对比，结果表明以结构面网络模拟结果为基础，应用模糊信息优化技术进行岩体质量评价能够取得很好的效果，解决了工程岩体质量的定量评价问题；(2)首次通过对已有的现场大型岩体变形试验结果的分析及相应试验点岩体结构分形特征的研究，建立了结构面分布的分形维数和岩体变形指标间的相关关系，并首次应用这种相关关系根据结构面网络模拟及分形维数计算结果确定了缺少试验结果的地下厂房各工程部位的岩体变形指标而且应用于围岩稳定性计算，解决了工程岩体宏观变形参数的评价问题；(3)应用在结构面模拟网络图上模拟岩体剪切破坏带的方法，得出了岩体在不同方向上的强度参数，结果表明岩体的强度特性与结构面的发育方向和程度有着明显的关系，各个岩体结构均质区具有不同的强度参数；同时还通过应用拉格朗日差分法对岩体强度指标的数值分析并与按连通率加权平均计算法对比表明，按连通率加权平均计算法在考虑合理的折减系数情况下，可以简单快速地对缺少现场大型试验结果的具体工程部位实现岩体强度指标的估计，通过以上研究解决了工程岩体抗剪强度参数的评价问题；(4)通过结构面网络分形维数的研究，应用分形损伤耦合分析方法，研究了厂房区岩体的损伤特征，并首次引入"损伤度"的概念，建立了应用最大主损伤评价岩体质量的指标体系。

Who? I never heard of him, Paul said, before asking teammate James Posey if he had heard of him.

赛后，科比说，他一直都是一名非常出色的射手，今天他打得很棒。

When I joined the company, I rotated around the different sections.

我加入这个公司时，轮换过几个不同的部门。