表位

It is of great importance to find out the antigenic epitope of TRP-2 as an antigen in pigmental cells, hi order to set up a foundation to investigate the epitope of TRP-2, we studied on the cloning and expression of human TRP -2 code gene.

确定其抗原表位对于深入研究该抗原及临床应用具有极其重要的意义，为了给TRP—2抗原表位确定的研究打下基础，本研究克隆了人TRP—2编码基因并表达了TRP—2蛋白。

The results showed that the purified serum can recognize MUC1 protein ..proliferation assays of T-cells from spleen of Balb/C inoculated with the positive phages showed that T-cells were markely proliferated in absence of adjuvant during immunization , which demenstrated that phage might be a proper vector for the immunization.

为证实模拟抗原的免疫原性，进一步用展示MUC1抗原模拟表位的噬菌体阳性克隆免疫小鼠，3次免疫小鼠后，血清(1：1000)中能检测到较强的MUC1抗体阳性；以最大OD450值之纯化血清做为一抗行蛋白印迹分析，结果显示：纯化血清可识别MUC1蛋白；小鼠脾脏淋巴细胞增殖率检测结果表明：噬菌体展示肽表位可在无佐剂存在情况下，大大促进小鼠的脾淋巴细胞的增殖，证明噬菌体是一较好的生物载体。

Objective To evaluate the potential application of the recombinant multiple epitope antigen in diagnosis of toxoplasmosis.

目的 评价弓形虫多表位基因重组抗原在弓形虫感染诊断中的效果，探索多表位抗原在弓形虫免疫检测中的应用前景。

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。

Nucleoprotein is also a major immunogenetic antigen for SARS; however, several tests have shown that N protein had the reactivity with other coronavirus positive sera. It is therefore an advantage of S protein as recombinant target antigen for SARS serum diagnosis compared with recombinant N protein. Moreover, S protein has been shown to be the major antigenic determinant that induces neutralizing antibodies and protective immunity to SARS-CoV, so it is also important in designing SARS genetic engineering vaccine.

SARS-CoV S 蛋白由1255 个氨基酸组成，体外表达如此大的蛋白很困难，而且表达蛋白的可溶性不高；其次，实验已经证实，全长S 蛋白含有大量无关的表位，致使表达的全长S蛋白对SARS 阳性血清的检出率低于S 蛋白片段；另外作为疫苗研究，S 蛋白上大量无关表位作为异体蛋白进入体内，会引发机体的过度免疫反应，这也是SARS-CoV 可能的致病机理之一。

The mimic antigen of paragonimus had definite diagnostic value in the paragonimiasis,and the five epitopes may mimic the natural antigen in the special structure.

结论肺吸虫模拟抗原对肺吸虫病具有一定的诊断价值，这5种表位可能从空间结构上模拟了天然抗原表位。

Results showed that methods,shows a wide application future in the analysis of proteinic epitope.

新的蛋白质抗原表位分析技术弥补了传统分析技术的不足，在蛋白质抗原表位分析中具有广阔的应用景。

Methods HBcAg specific CD8+ cells in whole blood samples of chronic hepatitis B patients were stained with soluble HLA-A2—HBV core 18-27 peptide tetrameric complex counted by flow cytometry.

应用人类主要组织相容性复合体A2的胞外段与HBcAg表位肽Tc18-27构建的四聚体结合流式细胞术检测人外周全血中该表位肽特异的CD8+CTL的数量，并以其占CD8+细胞总数的百分比表示。

Our study demonstrated that the epitope-specific CTLs function was defected. Interestingly, in contrast to previous study, our result found that direct ICCS assys using peptide core 18-27 were able to visulize a population of IFN-γin the absence of specific pentamer binding in patients.

值得注意的是，我们发现在慢性乙肝患者外周血中Treg频率明显增高且与表位特异性CTL的频率呈现负相关，这种现象给了我们新的提示，为此我们用磁珠分选出Treg后在体外作不同细胞亚群共孵育，从而研究Treg与表位特异性CTL的相互作用机制。

In this research, B2 gene of HCV was highly expressed as a fusion protein (Trx-B2) by cloning into pThioHisA, and induced specific anti-HCV antibody in higher titer than unmodified B2 protein after immunize mice or rabbit.The results showed antigen Trx-B2 has obvious immunogenicity and induced specific antibody,which might be able to serve as diagnosis HCV. Consequently, this PcAb of rabbit was used to coated Immulon plate, and a method of both antibodys sandwiching antigens was developed to detect 32 HCV positive serum and 32 nomal serum.Results show that ratio of positive are 87.5%and 43.75% respectively.

但缺点是该抗原所诱导的抗体滴度较低，可能是由于其分子量较小，所以本课题将人工合成的HCV复合多表位抗原基因B_2克隆到表达载体pThioHisA中，利用硫氧还蛋白融合方式对该抗原进行修饰，并在大肠杆菌中表达融合蛋白(Trx-B_2)，纯化该蛋白后免疫小鼠及家兔，结果表明用硫氧还蛋白修饰后的融合蛋白免疫小鼠和家兔后，得到的抗体滴度均高于未经修饰过的复合多表位抗原合成肽所诱导的抗体滴度，并从免疫后的兔血清中提取IgG包被酶联板，利用双抗夹心法分别检测了32份HCV阳性血清和32份正常人的血清，结果阳性率分别为87.5%和43.75%，符合率为71.88%。

However, as the name（read－only memory）implies, CD disks cannot be written onorchanged in any way.

然而，正如其名字所指出的那样，CD盘不能写，也不能用任何方式改变其内容。

Galvanizes steel pallet is mainly export which suits standard packing of European Union, the North America. galvanizes steel pallet is suitable to heavy rack. Pallet surface can design plate type, corrugated and the gap form, satisfies the different requirements.

镀锌钢托盘多用于出口，替代木托盘，免薰蒸，符合欧盟、北美各国对出口货物包装材料的法令要求；喷涂钢托盘适用于重载上货架之用，托盘表面根据需要制作成平板状、波纹状及间隔形式，满足不同的使用要求。