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11.
Dina Zand Jeffrey Hammer Robert Gould Richard Quarles 《Journal of neurochemistry》1991,57(3):1076-1079
The major 24- and 28-kDa glycoproteins in shark PNS and CNS myelin express high levels of the adhesion-associated HNK-1/L2 carbohydrate epitope. The 28-kDa protein, but not the 24-kDa protein, cross-reacts strongly with one of two anti-bovine P0 antisera not previously tested against fish myelin proteins. Shark PNS and CNS myelin also contains smaller amounts of high-molecular-weight HNK-1-positive proteins, including a prominent broad band in the 65-85-kDa range. Although myelin-associated glycoprotein (MAG) is well known to react with HNK-1 in some mammals, monoclonal and polyclonal anti-MAG antibodies did not react with the high-molecular-weight HNK-1-positive material in shark myelin, a result suggesting that it is not a MAG-like protein. The high expression of the HNK-1/L2 epitope in glycoproteins of shark myelin, including the major P0-related ones, suggests that this adhesion-related carbohydrate structure may have had an important role in the molecular evolution of the myelinating process. 相似文献
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SARS病毒M蛋白的二级结构和B细胞表位预测 总被引:4,自引:0,他引:4
以SARS病毒基因组序列为基础,采用GarnierRobson方法、ChouFasman方法和KarplusSchulz方法预测蛋白质的二级结构;按KyteDoolittle方案、Emini方案和JamesonWolf方案预测SARS病毒M蛋白的B细胞表位。预测结果表明,在SARS病毒M蛋白N端第11~20、27~36区段和第133~141区段可能是α螺旋中心;M蛋白分子N端第20~27、34~37,44~56,61~64,70~76,79~97,117~132,142~147,165~176区段和第216~221区段可能是β折叠中心。在M蛋白N端第5~6、40~44、105~107、112~116、189~190、202~203区段和第210~215区段具有较柔软的结构,有可能进行一定幅度的摆动或折叠而形成较复杂的三级结构。SARS病毒M蛋白N端第1~15、37~47、99~120、181~192区段和第196~215区段内或附近很可能是B细胞表位优势区域。以蛋白质的二级结构预测作为辅助手段,用抗原指数,亲水性参数和可及性参数预测SARS冠状病毒M蛋白的B细胞表位,为实验确定SARS病毒M蛋白的B细胞表位和免疫识别研究奠定了基础 。 相似文献
14.
同尾酶技术在构建疟疾多价重组DNA疫苗中的应用 总被引:6,自引:0,他引:6
同尾酶是一类识别不同核苷酸序列但能酶切产生相同粘性末端的限制性内切酶,依靠同尾酶的这种特性,可以根据需要将不同的DNA 片段进行灵活组合,获得各种排列顺序的多价表位重组疫苗.将这种方法用于疟疾多价重组DNA 疫苗的研制;BALB/c 小鼠免疫实验对所得重组疫苗PU286的免疫原性进行了测定. 相似文献
15.
抗禽流感病毒多表位DNA疫苗的构建及其免疫效力研究 总被引:17,自引:1,他引:17
多表位DNA疫苗是建立在常规DNA疫苗基础上的一种新型疫苗。它是用表位作免疫原,这样就比较容易在一个表达载体上克隆病原体的多个抗原基因中具有免疫活性的部分。本试验以H5N1亚型禽流感病毒的HA和NP基因及其表位为基础构建了4个重组质粒:1 pIRES/HA(表达全长的HA基因);2 pIRES/tHA(只表达HA基因的主要抗原表位区);3 pIRES/tHANpep(融合表达HA基因的抗原表位区和NP基因的3个CTL表位);4 pIRES/tHANpep-IFN-γ(用鸡的IFN-γ基因取代质粒pIRES/tHANpep中的neo基因)。分别用这4个重组质粒和空载体质粒pIRES1neo肌注免疫30日龄SPF鸡。免疫3次,间隔为2周,每次每只鸡的剂量为200μg。第3次免疫后两周以高致病性禽流感病毒H5N1强毒攻击,免疫及攻毒前后均采血检测HI抗体效价和外周血CD4+、CD8+T细胞的变化。结果发现,攻毒前各质粒免疫组均检测不到HI抗体,攻毒后1周存活鸡HI抗体效价迅速升高到64~256。流式细胞仪检测显示外周血CD4+、CD8+T细胞在疫苗免疫后都有不同程度的升高。空载体质粒对照组鸡(10只)在攻毒后3~8 d内全部死亡,其他各重组质粒免疫组鸡都获得了部分保护,保护率分别是:pIRES/HA组为545%(6/11),pIRES/tHA组为30%(3/10),pIRES/tHANPep组为36.3%(4/11), pIRES/tHANPepIFNγ组为50%(5/10)。这些结果表明我们构建的多表位DNA疫苗能够诱导机体产生特异性免疫应答,并在同型禽流感强毒攻击时对鸡只提供了一定的保护。 相似文献
16.
The Development of Multi-epitope Vaccines: Epitope Identification, Vaccine Design and Clinical Evaluation 总被引:5,自引:0,他引:5
A. Sette B. Livingston D. McKinney E. Appella J. Fikes J. Sidney M. Newman R. Chesnut 《Biologicals》2001,29(3-4):271-276
We have developed efficient methods for epitope identification and vaccine design. Our process for epitope selection based on the combined use of motif analyses, binding assays and immunogenicity evaluations is described. We also describe how the projected population coverage and vaccine design can be optimized. Finally, it is discussed how vaccine potency is evaluated by immunogenicity and antigenicity assays. 相似文献
17.
信号肽和辅助性T细胞表位增强HBV核心抗原DNA疫苗诱导的免疫应答 总被引:13,自引:1,他引:13
为增强HBVDNA疫苗的免疫效率 ,于HBV核心抗原 (HBcAg)基因 5′末端引入人IL 2信号肽和一个通用型辅助性T淋巴细胞表位基因 ,并构建成DNA疫苗 ,转染COS7细胞后经ELISA检测出分泌型HBcAg。通过肌肉注射途径分别将这种DNA疫苗和编码天然HBcAg的DNA疫苗免疫BALB/c小鼠 ,检测小鼠的血清抗体、T细胞增殖和细胞毒性T淋巴细胞反应 ,结果表明前者诱导细胞和体液免疫应答的强度均明显超过后者 ,且更趋向于T辅助细胞 1(Th1)型免疫应答 ,故其对慢性HBV感染的治疗可能有潜在的应用价值 相似文献
18.
Wangxiao He Pietro Mazzuca Weirong Yuan Kristen Varney Antonella Bugatti Alfredo Cagnotto Cinzia Giagulli Marco Rusnati Stefania Marsico Luisa Diomede Mario Salmona Arnaldo Caruso Wuyuan Lu Francesca Caccuri 《Biochimica et Biophysica Acta (BBA)/General Subjects》2019,1863(1):13-24
Background
HIV-1 matrix protein p17 variants (vp17s) detected in HIV-1-infected patients with non-Hodgkin's lymphoma (HIV-NHL) display, differently from the wild-type protein (refp17), B cell growth-promoting activity. Biophysical analysis revealed that vp17s are destabilized as compared to refp17, motivating us to explore structure-function relationships.Methods
We used: biophysical techniques (circular dichroism (CD), nuclear magnetic resonance (NMR) and thermal/GuHCL denaturation) to study protein conformation and stability; Surface plasmon resonance (SPR) to study interactions; Western blot to investigate signaling pathways; and Colony Formation and Soft Agar assays to study B cell proliferation and clonogenicity.Results
By forcing the formation of a disulfide bridge between Cys residues at positions 57 and 87 we obtained a destabilized p17 capable of promoting B cell proliferation. This finding prompted us to dissect refp17 to identify the functional epitope. A synthetic peptide (F1) spanning from amino acid (aa) 2 to 21 was found to activate Akt and promote B cell proliferation and clonogenicity. Three positively charged aa (Arg15, Lys18 and Arg20) proved critical for sustaining the proliferative activity of both F1 and HIV-NHL-derived vp17s. Lack of any interaction of F1 with the known refp17 receptors suggests an alternate one involved in cell proliferation.Conclusions
The molecular reasons for the proliferative activity of vp17s, compared to refp17, relies on the exposure of a functional epitope capable of activating Akt.General significance
Our findings pave the way for identifying the receptor(s) responsible for B cell proliferation and offer new opportunities to identify novel treatment strategies in combating HIV-related NHL. 相似文献19.
Unterreitmeier S Fuchs A Schäffler T Heym RG Frishman D Langosch D 《Journal of molecular biology》2007,374(3):705-718
Interactions of transmembrane helices play a crucial role in the folding and oligomerisation of integral membrane proteins. In order to uncover novel sequence motifs mediating these interactions, we randomised one face of a transmembrane helix with a set of non-polar or moderately polar amino acids. Those sequences capable of self-interaction upon integration into bacterial inner membranes were selected by means of the ToxR/POSSYCCAT system. A comparison between low/medium-affinity and high-affinity sequences reveals that high-affinity sequences are strongly enriched in phenylalanine residues that are frequently observed at the − 3 position of GxxxG motifs, thus yielding FxxGxxxG motifs. Mutation of Phe or GxxxG in selected sequences significantly reduces self-interaction of the transmembrane domains without affecting their efficiency of membrane integration. Conversely, grafting FxxGxxxG onto unrelated transmembrane domains strongly enhances their interaction. Further, we find that FxxGxxxG is significantly over-represented in transmembrane domains of bitopic membrane proteins. The same motif contributes to self-interaction of the vesicular stomatitis virus G protein transmembrane domain. We conclude that Phe stabilises membrane-spanning GxxxG motifs. This is one example of how the role of certain side-chains in helix-helix interfaces is modulated by sequence context. 相似文献
20.