查询词典 DNA

The first reaction mixture consisted of 5uL 10×PCR buffer,0.5uL 20mmol/L dNTP,1uL of primer F1、R2(25umol/L),0.5uL Taq (2.5U),FTA filtering,42μL of double-distilled water,the whole system was 50μL;The reaction was run under the following conditions:DNA pre-denaturation at 94℃for 5 min;DNA denaturation at 94℃for 30 s,primer annealing at 58.8℃for 1min, extension at 72℃for 30s,for 25 cycles,and extension at 72℃for 5min.

第一轮适宜反应体系分别为：5uL的10×PCR buffer，0.5uL的20mmol/LdNTP，引物F1、R2各1uL(25umol/L)，0.5uL的Taq酶(2.5U)，含有基因组DNA模板的滤膜，用去离子水补足到50 uL。

Nine different aphid-resistant alfalfas and four cross bred F1 were used in this experiment to investigate better dosage in SSR technique system and to seek the best combination among factors.At last,the best SSR technique system was established. The Taq DNA polymerase was 0.2μL(5U/μL),the 10×Buffer(Mg2+Plus)was 2.5μL , the dNTP Mixture was 2μL ( 2.5mmol/L), the primer was 1.5μL (10μmol/L),the template DNA was 1μL,the ddH2O was 16.3μL ,and the whole system was 25μL.

以九个不同抗蚜级别的苜蓿品种及四个杂交F1代为试材，摸索SSR反应体系中各个影响因素的浓度和用量，确定适合苜蓿抗蚜虫基因定位的SSR反应体系：其中Taq DNA聚合酶0.2μL(5U/μL)，10×Buffer(Mg2+Plus) 2.5μL，dNTP Mixture 2μL (各2.5mM)，引物各1.5μL (10μM)，模板DNA1μL，ddH2O 16.3μL补足25μL。

In order to develop a new path for gene mapping of fish, the digoxigenin and tritium labeled in situ hybridization techniques combined with multiple banding analysis on the pachytene bivalents of M. albus were studied using the wheat ribosomal RNA genes, pig growth hormone gene and MaSoxl (M. albus SRY boxl) DNA segment as the probes. Moreover, the techniques of random primed in situ DNA synthesis were also developed on the bivalents of M.

以两种异源基因和Ma Soxl为探针，采用地高辛或氟标记的原位杂交技术，结合点渍法和DNA印渍法分析，率先在黄鳝减数分裂二价体上开展基因定位研究，并与多重带带型分析相结合，以期为构建黄鳝染色体框架图奠定初步基础，为迅速改变鱼类基因定位研究的落后局面而开辟新的途径。

Methods The DNA chip was fabricated,on which oligonucleotide bougies′length was 19 base pairs,and the concentration was above 15 μmog/L.

方法自制DNA芯片，每条寡核苷酸探针片段长度为19bp，点样浓度在15μmol/L，以痰液为待检标本，分别用不同引物、Mg2+、核苷酸、DNA聚合酶浓度等进行多重聚合酶链反应；分别在不同时间、温度下进行杂交，分别在不同酶作用时间和显色时间进行分析，比较不同条件芯片检测结果。

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。

Owing to the linear relation of the quenching between carbaryl and ct DNA, carbaryl is regarded as one kind of fluorescent probe to detect DNA.

甲萘威与小牛胸腺DNA之间的猝灭具有良好的线性关系，所以甲萘威可以用于荧光探针而对脱氧核糖核酸进行定量分析。

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。

The internal transcribed spacer （ITS, contains ITS-1, 5.8S nuclear ribosomal DNA, ITS-2） and 28S nuclear ribosomal DNA-LSU were amplified by PCR, sequenced and analyzed by Chromas and DNASTAR softwares, and the RNA secondary structure of 28S rDNA-LSU was analyzed by DNAMAN software.

抽提成虫基因组DNA，扩增其ITS（包括ITS-1、5.8S rDNA和ITS-2）和28S rDNA-LSU序列，测序并分析以上序列，以及28S rDNA-LSU序列RNA二级结构。

In aid of the domain flexibility and requirement of cofactors, POU proteins exhibit very complicated ability to bind and recognize DNA in function of regulator and transcription factors.

两个亚结构域与DNA相互作用，在连接区可塑性和辅因子的帮助下，POU蛋白作为调控因子和转录因子，显示出错综复杂的DNA结合和识别能力。

I can not make it blossom and suits me

我不能让树为我开花

When temperatures are above approximately 80 °C discolouration of the raceways or rolling elements is a frequent feature.

当温度高于 80 °C 左右时，滚道或滚动元件褪色是很常见的特征。