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聚合酶

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Target DNA fragments were amplified by polymerase chain reaction from the liver tissue of Columba livia using a pair of specific primers, and the amplified PCR products were cloned into pMD18-T vector.

利用特异引物,通过聚合酶链式反应(polymerase chain reaction, PCR)技术,从家鸽肝脏组织的总DNA中扩增到目的片段,并将扩增产物克隆到pMD18-T载体中,经菌落PCR与酶切鉴定、序列测定及序列分析。

Methods: A polymerase chain reaction assay with oligonucleotide primers homologous to a portion of the urease C and cytotoxin associated gene A of Hp was used to test the presense of Hp in dental plaque.

依据特异的尿素酶C 基因和 cag A基因设计引物,建立聚合酶链反应方法,检测65例慢性胃病患者不同牙齿的龈上和龈下菌斑中的Hp.6例胃Hp阳性的胃炎患者,经药物治疗后,检测口腔Hp。

In order to understand insertion/delation polymorphism of the angiotensin-converting enzyme gene in pilots,and to explore the relationship between ACE gene I/D polymorphism and the perfomance of the pilots,the polymerase chain reaction was used to determine the genotypes for an I/D polymorphism in intron 16 of the ACE gene in 118 pilots and 96 healthy subjects as controls.

为了解飞行员血管紧张素转换酶基因插入或缺失多态性情况,探讨ACE基因多态性与飞行员耐力可能的关系,用聚合酶链反应扩增技术检测118例飞行员和96例健康对照者的ACE基因I/D多态性。

Objective To compare the efficiency of constructing three truncated Echinococcus multilocularis18 gene expression vector using two clone methods.

目的比较3种截短的泡球蚴Em18基因聚合酶链反应扩增产物经TA克隆和直接酶切2种方法,在原核表达质粒构建中的运用。

The LV messenger ribonucleic acid levels of contractile and calcium regulatory genes were measured by quantitative real time polymerase chain reaction and normalized for glyceraldehyde 3-phosphate dehydrogenase.

采用定量实时聚合酶链反应测定心室收缩蛋白和钙调蛋白基因信使RNA水平,采用3-磷酸甘油醛脱氢酶标准化。

Methods BALB/c mice were anesthetized with ether and infected intracranially with 20 μl 4.5×108 plaque forming units of coxsackievirus B3. CVB3 RNA in mice brain was detected by reversal transcription polymerase chain reaction and the pathological changes of brain tissues were observed by HE staining.

BALB/c纯系小鼠分两组,实验组小鼠颅内直接注射滴度为4.5×108 pfu的柯萨奇病毒B3悬液20 μl,对照组颅内注射生理盐水20 μl,应用逆转录酶聚合酶链反应法测定脑组织中CVB3 RNA含量,HE染色观察脑组织病理学的改变。

To prove that the cloned DNA fragment can express tryptopanase,a new plasmid pET28C-TnaA , in which the cloned DNA fragment was located downstream of T7 promoter on pET28c was constructed and transformed into host BL21(DE3),a BL21 lysogen of bacteriophage DE3 in which the only promoter known to direct transcription of the T7 RNA polymerase gene is the lacuvS promoter ,which is inducible by IPTG.

为了证明质粒上的基因能表达出有活性的色氨酸酶,将这个DNA片段克隆到PET28c质粒的BamHI和HindⅢ位点上,使该片段受T7 RNA聚合酶的启动子控制,然后转化噬菌体DE3的溶源菌BL21(DE3)。

Methods The effect of ginsenoside-Ro on murine splenocytes proliferation was studied using [3H] thymidine incorporation assay. Effects of ginsenodide-Ro on the production of cyrokines interleukin-2 (IL-2), interferon-γ and interleukin-4 (IL-4) from mutine splenocytes were detected by ELISA method. Effects of ginsenoside-Ro on mRNA level of TH1 cytokine IFN-γ and Th2 cytokine IL-4 were evaluated by reverse transcription polymerase chain reaction analysis.

方法[3H] TdR参人法检测人参皂有-Ro对小鼠脾淋巴细胞增殖的影响;酶联免疫吸附法检测人参皂苷-Ro对小鼠脾淋巴细胞产生细胞因子白介素-2、干扰素-γ和白介素-4的影响;逆转录聚合酶链式反应分析法研究人参皂苷-Ro对小鼠脾淋巴细胞引干扰素-Υ、白介素-4mRNA表达的影响。

METHODS: A total of 264 chronic HBVinfected patients were enrolled in our study, and were divided into lowlevel HBsAg group(147 cases) and highlevel HBsAg group(117 cases) based on the HBsAg level or were divided into immune tolerance stage, immune clearance stage and nonactive stage based on the natural history. Realtime PCR and microparticle enzyme immunoassay were used to determine the content of HBV DNA and HBV M in serum samples of highlevel and lowlevel HBsAg patients, respectively, then the quantitative results were compared.

对264例慢性HBV感染者的HBsAg浓度(高浓度HBsAg组、低浓度HBsAg组)及自然史(免疫耐受期、免疫清除期、非活动期)进行分组,采用实时荧光聚合酶链反应和微粒子酶免疫试验对147例低浓度HBsA组和117例高浓度HBsAg组的血清标本进行HBV DNA和HBV M的定量测定并进行比较。

Methods The total RNA was extracted from the brain of human embryo. The segment of open read frame of DOC-1 was amplified by RT-PCR and was further cloned into the vector, pMD18-T, and the prokaryotic expression vector, pGEX-4T-1, by turns, to produce the new construct pGEX-4T-1-DOC-1. After restriction enzymes digestion analysis and sequenced, pGEX-4T-1-DOC-1 was transformed into E.coli BL21(DE3). GST-p12 recombinant protein was expressed under IPTG induction and purified by affinity column.

从人胎脑组织中提取总RNA,经逆转录-聚合酶链式反应扩增DOC-1的CDS序列,再通过基因重组技术将该基因片段依次克隆到pMD18-T和pGEX-4T-1载体中,构建融合表达载体pGEX-4T-1-DOC-1,经酶切、测序鉴定后,用该重组质粒转化E.coliBL21,用IPTG诱导表达,Glutathione Sepharose 4B柱亲和层析纯化重组蛋白。

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