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从志贺氏1型痢疾村菌LPS中分离纯化出料捩的,以ADH为连接剂将其与TT结合形成O-SP-TT结合疫苗,并用此结合疫苗免疫NIH小鼠,结果显示使用O-SP免疫后,小鼠血清中没有抗LPS抗体产生,而用L-SP-TT免疫后鼠血清中生了抗LPSIgG和IgM抗体,且IgG抗体水平高于IgM抗体;O-SP-TT免疫组等二次和第三次免疫后IgG我有显著的升高(P〈0.01),但第二次和第三次免疫后血清IgM 相似文献
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Gag和Env蛋白是人I型免疫缺陷病毒(Human immunodeticiency virus type 1,HIV-1)的结构蛋白,是HIV-1诱导机体产生体液免疫和细胞免疫的主要抗原。本实验通过多交亚克隆,将env基因以正确的三联密码读框插入gag基因的下游制备了HIV-1 gag-env嵌合基因,并将嵌合基因分别置于痘苗病毒p7.5启动子和牛痘病毒A型包涵体(ATI)启动子的下游,经过同源 相似文献
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为探讨HCV/HBV 复合疫苗的可行性,将合成的丙型肝炎病毒(HCV)复合多表位抗原基因PCX与HBsAg 基因连接成PCXS基因,与β-半乳糖苷酶(GZ)基因融合后在大肠杆菌及减毒鼠伤寒沙门氏菌中获得表达.目的蛋白GZ-PCXS可被抗-HBs 及抗-HCV 抗体所特异识别.GZ-PCXS抗原皮下注射免疫ICR小鼠后,诱发了较高水平的抗-GZ-PCXSIgG反应.构建的重组减毒鼠伤寒沙门氏菌SL3261(pWR/PCXS)口服免疫小鼠后,诱发了高水平的CD8+ T细胞增殖反应及抗GZ-PCXSIgG反应.所有免疫小鼠均未见明显的毒副作用.该研究揭示,HCV/HBV 复合抗原可诱发特异性体液免疫及细胞免疫应答,而活菌苗口服可能是理想的免疫途径,为HCV/HBV 双价疫苗研究提供了一定的理论及实验依据. 相似文献
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将编码丙型肝炎病毒(HCV)E2蛋白417~750位氨基酸的DNA片段 克隆到真核表达载体pcDNA 3.1(-)中的CMV IE启动子下游,构建成HCV E2重组真核表达质粒 pcE2。ELISA法检测pcE2 DNA免疫兔血清中的E2抗体变化和维持规律,结果显示免疫20d已有 抗体产生,30d后开始进入高峰,40d时达到最高值,至第90d抗体水平保持平稳,抗体滴度 达到1∶1600左右。流式细胞计数仪(FACS)检测pcE2 DNA免疫鼠CD4+、CD8+T淋巴细胞变 化情况,与注射空载体pCDNA3.1(-)的阴性鼠相比,CD4+淋巴细胞水平略有上升,CD8+ 细胞水平有较大升高,增幅达35.46%。免疫组化检测结果显示注射pcE2的小鼠组织中有明显 的阳性着色,而注射pcDNA3.1(-)的对照组小鼠免疫组化结果为阴性。以上结果表明:pcE2 在实验动物内表达出的HCV E2蛋白可以引起免疫动物的体液免疫应答和细胞免疫应答,尤其 是MHC-1限制性杀伤性CD8+T淋巴细胞水平的提高对清除 病毒是十分有利的,因此HCV E2 DNA免疫有可能成为预防和治疗HCV感染的一条新途径。 相似文献
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用丙肝病毒C+E1区真核表达质粒pcDNA-HCV/C+E1按400ug/只剂量免疫BALB/C小鼠,14周后同一剂量再加强免疫一次。加强免疫后2周,在50%PEG1450介导下将脾细胞与SP2/0小鼠骨髓瘤细胞(51)融合。实验结果融合率达54.3%(313/576)。阳性率为5.4%(17”313)。克隆化后得到6株稳定分泌抗丙肝病毒C区单克隆抗体杂交瘤细胞株。这6株杂交瘤均产生IgM抗体,接种BALB/C小鼠后产生腹水的效价为164-1320(ELISA)。 相似文献
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将间接ELISA、非放射性分子杂交和RT-PCR三种方法应用于水稻草矮病毒(RGSV)的检测。结果表明,利用自制的融合蛋白GST-NC的抗血清检测RGSV的灵敏度为1mg鲜重的病株叶片或84ng提纯病毒,利用地高辛(DIG)标记的DNA探针NC的点杂交方法检测RGSV的灵敏度为50μg病叶或6ng病毒,而RT-PCR的检测灵敏度则为10μg病叶或2ng的病毒,对上述三种方法的灵敏度和可操作性也进行 相似文献
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将系列缺失的HIV1长末端重复序列(LTR)和全长的gagORF置于痘苗病毒载体中,经同源重组和血球吸附试验,成功地构建了6株重组痘苗病毒。免疫印迹和免疫酶试验检测均表明,6株重组病毒的Gag蛋白表达量因LTR不同而有明显差异,表明HIV1的LTR及其下游基因置于痘病毒启动子控制下,在痘苗病毒中表达时有下述特点:(1)不同的痘苗病毒启动子与全长LTR相互作用,对gag基因表达有显著不同的调控效果;(2)NR序列对Gag蛋白表达没有明显影响;(3)EN序列不能被重组痘苗病毒表达系统识别;(4)TAR序列可提高Gag蛋白的表达量;(5)U5区及下游非翻译序列不影响Gag蛋白的表达。 相似文献
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我国首次发现的HIV—1和HIV—2双重感染样品中病毒部分基因的序列特 … 总被引:1,自引:0,他引:1
上海市卫生检疫局送检了一例HIV-1和HIV-2抗体检测均呈阳性的双重感染样品,对其感染的HIV前病毒的gag和env基因区进行了序列分析,首次阐明我国发现的HIV双重感染样品的HIV部分基因特征。从HIV感染者淋巴细胞(peripheral blood mononuclear cells,PBMC)中提取前病毒DNA,分别使用HIV-1和HIV-2特异性引特用套式PCR扩增HIV-1和HIV-2 相似文献
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It was shown by electron microscopic study that in the presence of inhibitor of myosin ATPase protein P55 the long fibrils of myosin became short. 相似文献
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Rabbit antiserum prepared against a cyclic 19-amino-acid peptide predicted from the sequence of the viral mos gene (v-mos) of Moloney murine sarcoma virus not only recognized v-mos gene products but also specifically detected a 55,000-Mr polypeptide expressed in a variety of cells that grow on solid surfaces. This normal cellular protein, previously shown to be reduced in amount in cells expressing the v-mos gene, was found to be the intermediate filament structural protein, vimentin. This conclusion was reached by comparing relative mobilities in denaturing gels, isoelectric points, immunoreactivities, location in the cell, and peptide maps. 相似文献
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E S Svanidze G A Stefanenko L S Grigor'eva N N Bokachadze M M Zaalishvili 《Biofizika》1981,26(5):926-928
A method of minor protein P55 isolation from extract of soluble proteins of A-zone of the sarcomere from rabbit skeletal muscle is described. It is shown that this protein inhibits Ca2+-ATPase of myosin and Mg2+-ATPase of reconstructed actomyosin, but it does not affect superprecipitation of actomyosin. The molecular weight which is determined by mobility and its polypeptide chain polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate is about 35 000 dalton. 相似文献
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Keetch CA Bromley EH McCammon MG Wang N Christodoulou J Robinson CV 《The Journal of biological chemistry》2005,280(50):41667-41674
Transthyretin is a tetrameric protein associated with the commonest form of systemic amyloid disease. Using isotopically labeled proteins and mass spectrometry, we compared subunit exchange in wild-type transthyretin with that of the variant associated with the most aggressive form of the disease, L55P. Wild-type subunit exchange occurs via both monomers and dimers, whereas exchange via dimers is the dominant mechanism for the L55P variant. Because patients with the L55P mutation are heterozygous, expressing both proteins simultaneously, we also analyzed the subunit exchange reaction between wild-type and L55P tetramers. We found that hybrid tetramers containing two or three L55P subunits dominate in the early stages of the reaction. Surprisingly, we also found that, in the presence of L55P transthyretin, the rate of dissociation of wild-type transthyretin is increased. This implies interactions between the two proteins that accelerate the formation of hybrid tetramers, a result with important implications for transthyretin amyloidosis. 相似文献
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《CMAJ》1971,104(11):1016-1017
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TNF与多种疾病密切相关。为了获得大量具有生物学活性的可溶性TNF受体用以拮抗TNF的毒性作用,在原核表达系统中表达了TNFR(P55)的胞外区与TrxA的融合蛋白。将TNFR(P55)胞外区去信号肽的前三个结构域基因克隆入融合蛋白表达载体pET-32a,在大肠杆菌BL21(DE3)中高效表达了TrxA-TNFR融合蛋白。表达产物以包涵体形式存在,经过变性和复性,并经镍金属鳌和柱亲和层析纯化,得到了纯度较高的可溶性受体蛋白的初纯品。免疫学实验及L929细胞体外实验均表明:该蛋白具有TNFR(P55)特异的抗原性、与TNF结合的活性以及良好的抑制TNF的TNF生物学活性。 相似文献
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Anders Öhman Tommy Öman Mikael Oliveberg 《Protein science : a publication of the Protein Society》2010,19(1):183-189
The ribosomal protein S6 from Thermus thermophilus has served as a model system for the study of protein folding, especially for understanding the effects of circular permutations of secondary structure elements. This study presents the structure of a permutant protein, the 96‐residue P54‐55, and the structure of its 101‐residue parent protein S6wt in solution. The data also characterizes the effects of circular permutation on the backbone dynamics of S6. Consistent with crystallographic data on S6wt, the overall solution structures of both P54‐55 and S6wt show a β‐sheet of four antiparallel β‐strands with two α‐helices packed on one side of the sheet. In clear contrast to the crystal data, however, the solution structure of S6wt reveals a disordered loop in the region between β‐strands 2 and 3 (Leu43‐Phe60) instead of a well‐ordered stretch and associated hydrophobic mini‐core observed in the crystal structure. Moreover, the data for P54‐55 show that the joined wild‐type N‐ and C‐terminals form a dynamically robust stretch with a hairpin structure that complies with the in silico design. Taken together, the results explain why the loop region of the S6wt structure is relatively insensitive to mutational perturbations, and why P54‐55 is more stable than S6wt: the permutant incision at Lys54‐Asp55 is energetically neutral by being located in an already disordered loop whereas the new hairpin between the wild‐type N‐ and C‐termini is stabilizing. 相似文献
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Jian-qiang Li Jun-jie Yang Xiu-juan Fan Zhen-peng Sun Yan Sun Huan Li Zi-xin Meng Wei Li 《中国病毒学》2012,27(1):10-18
Enterovirus 71 (EV71) is a member of the Entero-virus genus of the Picornaviridae family and is the major cause of Hand,foot,and mouth disease (HFMD) in children.Different strains from Gansu were clone... 相似文献