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We demonstrated that recombinant baculovirus vectors expressing p35 (Ac-CMV-p35) and eGFP could be transduced into human kidney 293 cells efficiently. The level of transgene expression was viral dose dependent and high-level expression of the target gene could be achieved under the heterogonous promoter.

本研究证明重组杆状病毒载体Ac-CMV-p35和Ac-CMV-GFP能有效地转导人胚肾293细胞,并介导外源基因高效表达,目的基因的表达水平与杆状病毒的感染剂量成正线性相关。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank,选择包含人主要T细胞抗原表位序列的人PSCA基因片段,应用异种化抗原设计技术,保留人T细胞抗原表位,设计异种化PSCA融合抗原片段;(2)根据核酸序列按中心模板法设计引物,应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因,以PCR、限制性酶切和DNA序列测定法进行鉴定:(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点,采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4),并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中,转染293T细胞,借助免疫荧光+流式细胞术考察插入片段表达效率,最终选定PSCA_3片段进行下一步研究;(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下,插入pVAX1-neo-IRES—GM/B7载体中,构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB,并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况;(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞,制备小鼠前列腺癌模型,并采用股四头肌肌肉注射+电脉冲法(Electroporation,EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB,同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照,通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率,来评价该DNA疫苗在小鼠体内的抑瘤效果。

A suitable expression system is required for heterologous expression of human lactoferrin.

目前常用的表达系统中,动物乳腺是人乳铁蛋白基因表达较为合适的器官。

It was shown that the resultant cybrid HL-R cells incorporated with reticulocyte cytoplasts became differentiated 〓 bytalterationcin growth parameters, which might be defined as decancerization:(1) loss of growth ability in soft agar medium;(2) nontumorigenic under heterotransplantation in their filial cellular generation to mude mice;(3) decrease of growth rate, cellular mitoxic index and DNA synthesis rate;(4) decrease or inhibition of c-myc gene expression;(5) expression of human globin gene products in 〓 subculture.

利用C-myc癌基因探针的分子杂交技术,未检测到胞质杂交细胞存在相当于C-myc基因特异的同源序列mRNA,提示杂交细胞癌基因表达严重受抑。

The expression of VEGFmRNA was valued by in situ hybridation method and the expression of Fit-1 and MV count was investigated by using immunohistochemical method.

应用原位分子杂交方法研究VEGFmRNA的表达,应用免疫组化方法研究Flt-1表达及MV的计数。结果:增生性瘢痕和瘢痕疙瘩组织中MV数目多於正常皮肤和扁平瘢痕组织中的MV数目。

Results compared with the normal villi,the quantity of P21 expression has no significant difference,the site of P21 expression in the hydatidiform mole is quite different.

结果 与正常绒毛相比,P21癌基因在葡萄胎组织中的表达量没有显著性差异,表达部位有明显不同。

Some factors, like expression stage in adenovirus and expression level in influenza, is also prominent in some viruses. In HPV and HIV, the hydropathy of the protein also plays its role, although not the main factor.

某些选择因素如腺病毒中基因的表达时相、流感病毒中基因的表达水平是比较明显地影响病毒密码子选择的因素,而在HPV和HIV基因所编码的蛋白的疏水性尽管对于病毒的密码子使用有影响,但不是最主要的因素。

In this study, the amount of ethylene and its key biosynthesis genes expression, typical allelochemical hydroxamic acid concentration and the key regulation genes (BX1 and BX9), as well as the dynamic expression and time of relevant defense key genes of secondary substance metabolism processes were compared in ethephon treatments of different concentration and duration.

为探明外源乙烯利对玉米化学防御作用诱导的浓度和时间效应,本文采用化学和分子生物学相结合的方法,研究了不同浓度乙烯利处理玉米后,叶片乙烯释放量及其合成调控关键基因ACS和ACO表达的变化,典型化感物质丁布的含量及其关键调控基因BX1和BX9表达的变化,以及其他调控次生代谢物的关键基因表达的时间动态。

To investigate the hyperglycemic effect on ICAM-1 expression of BBB, we took a dynamic observation of the expression on endothelium of BBB in SD rats with hyperglycemia for 3 days to 6 months, by immunohistochemical method.

为了解超负荷血糖条件下有否血脑屏障内皮细胞ICAM-1的表达,本研究采用SD大鼠尾静脉注射链脲霉素的方法,建立超负荷血糖模型。

The N gene ORF was then subcloned into pET-30a vector and the recombinant plasmid was transformed into E.coli BL21 (DE3) and induced with IPTG. The protein expression was determined by SDS-PAGE. The expressed protein had a molecular weight of 54.4 Ku that existed as inclusion body. Thin-layer scanning showed that the expression product accounted for 30.5% of the total bacterial proteins. The recombinant protein possessed native biological activity and could react with anti-PEDV hyperimmune serum in Western blot.

以阳性质粒为模板,用分别含有BamH Ⅰ和Xho Ⅰ酶切位点的上、下游引物扩增得到ORF,其PCR产物经BamH Ⅰ和Xho Ⅰ双酶切后定向克隆到pET-30a载体,构建的重组质粒命名为pET-30a-PN;将pET-30a-PN转化到大肠杆菌BL21 (DF3)中,在IPTG诱导下进行表达;SDS-PAGE结果表明表达出与预期大小相符的约54.4 Ku的重组蛋白,重组蛋白以包涵体形式存在;薄层扫描结果表明表达产物占菌体总蛋白的30.5%;Western blot分析表明表达的重组蛋白能与抗PEDV高免血清反应,说明该重组蛋白具有免疫学活性。

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When this condition occurs, inbound replication with the source partner is stopped on the destination domain controller and event ID 2042 is logged in the Directory Services event log.

计算机密码学是研究计算机信息加密、解密及其变换的科学,是数学和计算机的交义学科,也是一门新兴的学科。

Instructions: click on the thumbnails to see a larger image, then use the left-right arrow keys to scroll through the slideshow.

使用说明:滑鼠点在小图上即可放大观赏。开启后键盘左右键可用来换照片。

I can see it fastened to a nail next to the hole in the wall, but it is not fastened to that wire.

福尔摩斯说,我看到绳子是系在墙洞旁边的钉子上,而不是系在那根金属丝上。