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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。

Carthlicum, and was located on region of wheat chromosome 2AL near the gene Pm4b, it maybe either Pm4b, or a new gene linked to Pm4b. Temporarily, the gene was designated as PmPS5A. A dominant major gene conferring resistance to powdery mildew was identified in a BC〓F〓 population derived from a cross of amphidiploid Am9 and wheat cv. Laizhou953. The gene originated from T.

通过与含有已知抗病基因品种或品系的对该20个菌株反应模式比较和系谱分析,推测Am9/莱州953*〓 F〓含有Pm4b,波斯小麦PS5含有Pm4b与一个未知抗病基因组合;(DR147/Ae14)//莱州953*〓 F〓和硬粒小麦DR147含有Pm4a和一个未知抗病基因组合;尾状山羊草Ae14和小伞山羊草Y39含有新的抗白粉病基因。

Androgen receptor gene located on the X chromosome was first found as a susceptibility gene of AGA. Further more, the sex-determining gene located on the Y chromosome and the hairless gene on the autosome gene were also associated with AGA.

第一个发现的雄激素性秃发易感基因-雄激素受体基因,位于X染色体上;另外,位于Y染色体上的性别决定基因、常染色体上的脱发基因等也与雄激素性秃发发病有关。

Recently,many articles reported the close correlation between human height and gene polymor- phism,such as GH1 gene, VDR gene ,CYP19 gene,estrogen receptor gene and a few regions in the Y chromo- some.

GH1基因、 VDR基因、CYP19基因、雌激素受体基因以及Y染色体均与人的身材高矮有密切联系。

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推荐网络例句

The shaping method of noncircular part and the tool holder's radial motion characters in noncircular turning process are discussed in detail in the thesis.

论文详细研究了非圆零件的成型方法和加工过程中刀架的径向运动规律。

I have not really liked him,I do not like his this kind of disposition.

我没有真的喜欢他,我不喜欢他的这种性格。

As we know the price of traditional product is composed of the cost and the profit of the company involving market competition, monopolizes and many other factors.

我们知道作为传统的商品,定价的模式往往是在成本的基础上增加厂商的预计利润而形成其价格,当然也要考虑到市场竞争、垄断等其他方面的因素。