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We have recently determined the complete nucleotide sequence of the MS2 bacteriophage RNA molecule (Min Jou et al., 1972; Fiers et al., 1975,1976). Extensive segments of the sequence of the closely related phages R17 (23.9%) and f2 (11.5%) have been studied in other laboratories (Table 1). A comparison between the sequences of the phages MS2, R17 and f2 can now be given a functional significance by referring to the complete MS2 RNA sequence (Fig. 1).The estimation of the over-all degree of variation amounts to 3.9% (MS2-R17), 3.4% (MS2-f2) and 3.7% (R17-f2). All the differences observed can be accounted for by single base substitutions: transitions are highly predominant (86%). No change is found in the untranslated terminal regions and only two mutations occur in the intercistronic regions. From a total of 34 observed variable sites located in translated regions, 25 are neutral point mutations and only 9 lead to an (mostly rather conservative) amino acid change. The amount of variation in double-stranded regions (16 out of 36 cases) is much lower than the relative degree of secondary structure of the RNA (roughly two-thirds) would predict. Hence, there is clearly a selective pressure to preserve at least certain aspects of the three-dimensional conformation. 相似文献
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W Fiers S Neirynck T Deroo X Saelens W M Jou 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2001,356(1416):1961-1963
Soluble, recombinant forms of influenza A virus haemagglutinin and neuraminidase have been produced in cells of lower eukaryotes, and shown in a mouse model to induce complete protective immunity against a lethal virus challenge. Soluble neuraminidase, produced in a baculovirus system, consisted of tetramers, dimers and monomers. Only the tetramers were enzymatically active. The immunogenicity decreased very considerably in the order tetra > di > mono. Therefore, we fused the head part of the neuraminidase gene to a tetramerizing leucine zipper sequence; the resulting product was enzymatically active, tetrameric neuraminidase. The protective immunity induced by this engineered neuraminidase, however, remained fairly strain-specific. A third influenza A virus protein, the M2 protein, has only 23 amino acids exposed on the outer membrane surface. This extracellular part, M2e, has been remarkably conserved in all human influenza A strains since 1933. By fusing the M2e sequence to hepatitis B virus core protein, we could obtain highly immunogenic particles that induced complete, strain-independent, long-lasting protection in mice against a lethal viral challenge. Native M2 is a tetrameric protein and this conformation of the M2e part can also be mimicked by fusing this sequence to a tetramerizing leucine zipper. The potential of the resulting protein as a vaccine candidate remains to be evaluated. 相似文献
27.
A non-equilibrium thermodynamic model of oxidative phosphorylation is formulated, which allows us to take into account some non-local effects. In this way, we compute the influence of the tangential resistivity of the inner mitochondrial membrane to proton current, as well as that of the distance between active sites, on the stoichiometry and efficiency of energy conversion. 相似文献
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A universal influenza A vaccine based on the extracellular domain of the M2 protein. 总被引:27,自引:0,他引:27
The antigenic variation of influenza virus represents a major health problem. However, the extracellular domain of the minor, virus-coded M2 protein is nearly invariant in all influenza A strains. We genetically fused this M2 domain to the hepatitis B virus core (HBc) protein to create fusion gene coding for M2HBc; this gene was efficiently expressed in Escherichia coli. Intraperitoneal or intranasal administration of purified M2HBc particles to mice provided 90-100% protection against a lethal virus challenge. The protection was mediated by antibodies, as it was transferable by serum. The enhanced immunogenicity of the M2 extracellular domain exposed on HBc particles allows broad-spectrum, long-lasting protection against influenza A infections. 相似文献
29.
A G Bloor Y Jou C Hoeplinger J E Gartner G L Mayers R B Bankert 《Journal of immunology (Baltimore, Md. : 1950)》1982,128(3):1443-1449
In the course of this study, more than 3000 phthalate-specific antibody-forming cell hybrids were identified using the hybridoma technology. With the aid of a rapid screening assay and an extensive library of phthalate analogs, it was possible to assign selected hapten-specific clones to one of 11 distinct fine-specificity sets. This compartmentalization of the phthalate-specific hybridomas has made it possible to focus attention upon a single manageable portion of the phthalate-specific repertoire. Fourteen clones from a single fine-specificity set were selected for further immunochemical characterization. Five of these clones were found to secrete an antibody that was indistinguishable in isoelectric focusing. Affinity-purified, high-resolution anti-idiotype antibodies were prepared with specificity for the antibodies produced by one of these clones (i.e., 4C7). A major portion of the serologically defined private idiotype (4C7 IdI) was shown to be associated with the ligand-combining site. Our results indicate that the five clones that share a common spectrotype also express the 4C7 IdI. Two other independently derived clones from two distinct fusions also share this idiotype. The 4C7 IdI was also identified in affinity-purified anti-phthalate antibodies derived from a pool of phthalate-immune serum (conventional antibody) and from affinity-purified antibodies derived from a pool of serum from unimmunized BALB/c mice (natural antibody). The 4C7 IdI is thus considered to represent a repeating clonotype in the phthalate-specific repertoire of BALB/c mice, and will serve as one of several useful clonal markers that are being developed for studies of the mechanism regulating idiotype expression. 相似文献
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DNA pooling approach is a cost-saving strategy which is crucial for multiple-SNP association study and particularly for laboratories with limited budget. However, the biased allele frequency estimates cannot be completely abolished by κ correction. Using the SNaPshot™, we systematically examined the relations between actual minor allele frequencies (AMiAFs) levels and estimates obtained from the pooling process for all six types of SNPs. We applied principle of polynomial standard curves method (PSCM) to produce allele frequency estimates in pooled DNA samples and compared it with the κ method. The results showed that estimates derived from the PSCM were in general closer to AMiAFs than those from the κ method, particularly for C/G and G/T polymorphisms at the range of AMiAF between 20–40%. We demonstrated that applying PSCM in the SNaPshot™ platform is suitable for multiple-SNP association study using pooling strategy, due to its cost effectiveness and estimation accuracy. 相似文献