查询词典 sequencing problem

Methods All fecal swabs were freshly collected from outpatients with community-acquired diarrhea in our hospital and immediately plated onto six kinds of culture agars: XLD,TCBS,SS, CCDA, MacConkey agar and modified cefoperazone MacConkey agar. Phenotypic characteristics, 16S rRNA gene sequencing, transmission electronmicroscopyand drug susceptibility tests were employed to identify the isolated bacteria.

方法于2005年8月对杭州市第一人民医院肠道门诊就医患者采集粪样，分别接种SS、XLD、CCDA、TCBS、MacConkey琼脂以及改良头孢哌酮MacConkey琼脂等6种平皿，对分离到的可疑细菌进行常规生理生化实验、16S rRNA基因序列测定、透射电镜下形态观察及药敏试验。

In the study, RT-PCR and sequencing were employed to detect the expression levels of CCK-B receptor, CCKBRi4sv and gastrin mRNAs in 30 human gastric carcinoma and their corresponding normal tissues, 10 gastritis and 2 autopsied normal stomach specimens as well as in a gastric carcinoma cell line SGC-7901 cells.

本研究首先用RT-PCR及测序方法检测了30例人类胃癌组织及癌周正常胃粘膜、10例胃炎、2例尸检胃组织及胃癌细胞株SGC-7901中CCK-B受体、CCK-B受体的异位剪接变异体(intron 4-contained variant of cholecystokinin B receptor，CCKBRi4sv)及胃泌素基因的表达。

This programming includes the routing at branchings and the sequencing of goods flows at junctions.

这一个规画在分岐包括工作路线排定而且在联接包括货物的序列人流程。

Arabidopsis thaliana, a Cruciferous grass, is a model plant for molecular biology and has a close genetic relationship with brassicas. The genome sequencing was completed successfully al the end of 2000, which provided rich gene resource for biological research and application. Full use of these gene resources and searching the valuable genes for crop genetic improvement will become the targets of research.

拟南芥，是分子生物学研究的模式植物，与油菜同属于十字花科具有较近的亲缘关系。2000年底拟南芥全基因组测序工作的完成，为生物学的研究和应用提供了大量基因资源，充分挖掘这些基因资源以寻找有利用价值的基因应用于作物遗传改良将成为今后研究的重点。

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 objectives of this study are:(1) To amplify Tb wbbL gene encoding rhamnosyl transferase bypolymerase chain reaction from the genomic DNA ofM.tuberculosis H37RV strain;(2) To clone PCR product of Tb wbbLgene into a cloning vector pMD18-T for sequencing;(3) To subcloneTb wbbL gene to an expression vector pET16b to construct pET16b-Tb wbbL; and to overexpress Tb WbbL protein in E.coli BL21(DE3)under different induction conditions;(4) To establish theco-expression system for expressing chaperons of pKJE7 plasmidand soluble Tb WbbL protein in E.coli BL21(DE3) under differentinduction conditions;(5) To test expressed WbbL protein by SDS-PAGEand Western blot methods.

本论文的目的是：(1)利用 PCR 方法从结核分枝杆菌 H37Rv 菌株的基因组DNA 中扩增出 Tb wbbL 基因；(2)将 Tb wbbL 基因克隆到pMD18-T 克隆载体中，经 DNA 序列测定证实为正确的基因；(3)将 Tb wbbL 基因亚克隆到 pET16b 表达载体中并通过改变不同的诱导条件在大肠杆菌 BL21(DE3)中表达 Tb WbbL 蛋白质；(4)建立在 BL21(DE3)大肠杆菌中共表达 Tb WbbL 蛋白质与分子伴侣的体系，优化表达条件以高效表达出可溶性 Tb WbbL蛋白质；(5)用 Western blot 方法鉴定所表达的 Tb WbbL 蛋白质为 Tb wbbL 基因产物。

DNA sequencing confirmed that the constructs did have Chlamydial encoding genes in it which were correct in translation frame and sequence.

DNA 序列分析证实重组质粒含有相应的衣原体基因，其读码框架正确。

PCR with Family special primers was carried out to identify Family of PspA, and Clade of PspA was identified by means of bioinformatics according to the sequencing results.

通过家族专属引物PCR的方法鉴定这5种荚膜血清型肺炎球菌的PspA蛋白所属家族，并进一步根据基因测序结果通过生物信息学方法鉴定了所属支系。

PCR amplification, DNA sequencing and cladistic analysis.

PCR扩增、DNA序列测定、分支分析。

Chimborazo and Cotopaxi, took me by the hand.

越过琴博腊索山和科托帕克西山。

This car is in a good condition.

这辆车的状况很好。