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1.
The idiotypic network theory (N. K. Jerne, Ann. Immunol. 125, 373-389, 1974) predicts that any antibody that can be made by an individual would have its preexisting specific complementary B cells in its germline repertoire. We transplanted syngeneic BALB/c mice with live hybridoma cells and demonstrated the simultaneous presence of interacting idiotypic and anti-idiotypic B cells in an individual animal by immuno-cytoadherence assays. Furthermore, we demonstrate that interacting B cells displaying idiotypic and anti-idiotypic antibodies are subjected to lysis by complement. It is therefore tempting to speculate that this complement-sensitive interaction between idiotypic and complementary anti-idiotypic B cells in vivo may provide a mechanism for the regulation of B cell populations. 相似文献
2.
Intracellular localization of the viral polymerase proteins in cells infected with influenza virus and cells expressing PB1 protein from cloned cDNA. 总被引:19,自引:15,他引:4 下载免费PDF全文
The biosynthesis, nuclear transport, and formation of a complex among the influenza polymerase proteins were studied in influenza virus-infected MDBK cells by using monospecific antisera. To obtain these monospecific antisera, portions of cloned cDNAs encoding the individual polymerase proteins (PB1, PB2, or PA) of A/WSN/33 influenza virus were expressed as fusion proteins in Escherichia coli, and the purified fusion proteins were injected into rabbits. Studies using indirect immunofluorescence showed that early in the infectious cycle (4 h postinfection) of influenza virus, PB1 and PB2 are present mainly in the nucleus, whereas PA is predominantly present in the cytoplasm of the virus-infected cells. Later, at 6 to 8 h postinfection, all three polymerase proteins are apparent both in the cytoplasm as well as the nucleus. Radiolabeling and immunoprecipitation analyses showed that the three polymerase proteins remain physically associated as a complex in either the presence or the absence of ribonucleoproteins. In the cytoplasm, the majority of the polymerase proteins remain unassociated, whereas in the nucleus they are present as a complex of three polymerase proteins. To determine whether a polymerase protein is transported into the nucleus individually, PB1 was expressed from the cloned cDNA by using the simian virus 40 late promoter expression vector. PB1 alone, in the absence of the other polymerase proteins or the nucleoprotein, accumulates in the nucleus. This suggests that the formation of a complex with other viral protein(s) is not required for either nuclear transport or nuclear accumulation of PB1 protein and that the PB1 protein may contain an intrinsic signal(s) for nuclear transport. 相似文献
3.
Z L Hegedus H A Frank T I Steinman M D Altschule U Nayak 《Archives internationales de physiologie et de biochimie》1988,96(5):211-221
The fluorescence excitation and emission spectra observed in plasma from patients with chronic renal failure were reproduced by the generation of soluble lipofuscins in normal plasma samples by incubation with mixtures of L-dopa, dopamine, L-norepinephrine, L-epinephrine, 3-hydroxy-DL-kynurenine and 3-hydroxy-anthranilic acid for 24 h at 37 degrees C. Relative fluorescence intensity measurements consistently showed elevated plasma levels of the soluble lipofuscins in chronic renal failure: the means (n = 27) were 73.9 +/- 33.4 (SD) and 71.1 +/- 14.8 at emissions 413 nm and 445 nm respectively, in contrast to those of normal plasma samples (n = 11), 18.2 +/- 5.3 and 23.1 +/- 5.6. The maximum or shoulder at approximately 413 nm represents soluble lipofuscin that can be generated from 3-hydroxyanthranilic acid and the maximum or shoulder at approximately 445 nm represents soluble lipofuscins derived from the precursors listed above and probably from other related precursors. Gravimetric measurements also showed elevated levels of melanins in the plasma samples of patients with chronic renal failure: 2.72 +/- 0.38 mg/ml (n = 16), as compared to normal values: 1.70 +/- 0.10 mg/ml (n = 6). In individual patients haemodialysis reduced the fluorescence intensities to a range of 65-99% and the melanin levels to a range of 86-99% of the pre-dialysis values. 相似文献
4.
The high molecular weight aminoacyl-tRNA synthetase complex (the 24S complex) was isolated from rat liver by ultracentrifugation. The lysyl-tRNA synthetase (E.C. 6.1.1.6) was selectively dissociated by hydrophobic interaction chromatography on 1,6 diaminohexyl agarose followed by hydroxylapatite chromatography and DEAE chromatography. The lysyl-tRNA synthetase dissociated from the 24S synthetase complex was purified approximately to 2700 fold with 14% yield. 相似文献
5.
P. Nayak 《Journal of Phytopathology》1986,116(2):162-166
Twenty isolates of Xanthomonas campestris pv. oryzae (Ishiyama) Dye each from susceptible and resistant rice cultivars, were inoculated on the susceptible genotype IR-8 and resistant M Sungsong to measure the impact of host selection pressure on virulence change. Isolates virulent on resistant genotype tended to be less adaptive on susceptible cultivars. While isolates from susceptible genotype showed 2.2 times more general virulence (GV) than specific virulence (SV), isolates of resistant host origin had only 1.3 times GV, indicating that the resistant host plant displays considerable increase in virulence. The SV value increased 1.64 times after one cropping with resistants indicated the potential of the pathogen to change to be slow. 相似文献
6.
Complete nucleotide sequence of the nucleoprotein gene of influenza B virus. 总被引:7,自引:4,他引:3 下载免费PDF全文
A DNA copy of influenza B/Singapore/222/79 viral RNA segment 5, containing the gene coding for the nucleoprotein (NP), has been cloned in Escherichia coli plasmid pBR322, and its nucleotide sequence has been determined. The influenza B NP gene contains 1,839 nucleotides and codes for a protein of 560 amino acids with a molecular weight of 61,593. Comparison of the influenza B NP amino acid sequence with that of influenza A NP (A/PR/8/34) reveals 37% direct homology in the aligned regions, indicating a common ancestor. However, influenza B NP has an additional 50 amino acids at its N-terminal end. As is the case with influenza A NP, influenza B NP is a basic protein, with its charged residues relatively evenly distributed rather than clustered. The structural homology suggests functional similarity between the NP of influenza A and B viruses. 相似文献
7.
Influenza virus hemagglutinin expression is polarized in cells infected with recombinant SV40 viruses carrying cloned hemagglutinin DNA 总被引:35,自引:0,他引:35
Primary cell cultures of African Green monkey kidney (AGMK) contain polarized epithelial cells in which influenza virus matures predominantly at the apical surfaces above tight junctions. Influenza virus glycoproteins were found to be localized at the same membrane domain from which the virus budded. When polarized primary AGMK cells were infected with recombinant SV40 viruses containing DNA coding for either an influenza virus H1 or H2 subtype hemagglutinin (HA), the HA proteins were preferentially expressed at the apical surface in a manner identical to that observed in influenza virus-infected cells. Thus, cellular mechanisms for sorting membrane glycoproteins recognize some structural feature of the HA glycoprotein itself, and other viral proteins are not necessary for this process. 相似文献
8.
Characterization of Influenza Virus Ribonucleic Acid Duplex Produced by Annealing In Vitro 总被引:1,自引:1,他引:0 下载免费PDF全文
A ribonuclease-resistant ribonucleic acid (RNA) with a sedimentation coefficient of 12S was obtained by self-annealing influenza virus-specific RNA isolated from infected cells. It had the properties of double-stranded RNA. (i) Sedimentation behavior in sucrose gradient was independent of salt concentration. (ii) Thermal transition profile was sharp; the melting temperature is 83 C in 0.1 SSC (0.15 m NaCl plus 0.015 m sodium citrate) and 98 C in SSC. (iii) Buoyant density in cesium sulfate was 1.58 g/cm(3) compared to 1.64 g/cm(3) for single-stranded RNA. (iv) It gave rise to single-stranded RNA after denaturation. (v) The 12S RNA duplex contained both plus and minus strands of influenza virus. Labeled plus strands could be displaced by extraneous cold plus strands and extraneous (32)P-labeled plus strands could be incorporated into duplex after denaturation and reannealing. 相似文献
9.
R Nayak 《Biochemical and biophysical research communications》1992,184(1):467-470
5-Fluoro-2'-deoxyuridine is incorporated into DNA of mouse breast tumour in vivo. The incorporation is inhibited by thymidine. Part of the fluorodeoxyuridine is cleaved to fluorouracil and is incorporated into RNA. This incorporation is enhanced by thymidine. The result suggests that the major mechanism of action of the fluorouracil is due to its incorporation into RNA. 相似文献
10.
Influenza virus polymerase basic protein 1 interacts with influenza virus polymerase basic protein 2 at multiple sites. 总被引:2,自引:2,他引:0 下载免费PDF全文
Three polymerase proteins of influenza type A virus interact with each other to form the active polymerase complex. Polymerase basic protein 1 (PB1) can interact with PB2 in the presence or absence of polymerase acidic protein. In this study, we investigated the domains of PB1 involved in complex formation with PB2 in vivo, using coexpression and coimmunoprecipitation of the PB1-PB2 complex with monospecific antibodies. Results show that PB1 possesses at least two regions which can interact independently and form stable complexes with PB2. Both of these regions are located at the NH2 terminus of PB1; the COOH-terminal half of PB1 is not involved in interacting with PB2. Deletion analysis further demonstrated that the interacting regions of PB1 encompass amino acids (aa) 48 to 145 and aa 251 to 321. Linker insertions throughout the PB1 sequences did not affect complex formation with PB2. Deletion and linker-insertion mutants of PB1 were tested for polymerase activity in vivo. For this analysis, we developed a simplified assay for viral polymerase activity that uses a reporter chloramphenicol acetyltransferase gene containing the 5' and 3' ends of influenza viral promoter and nontranslating regions (minus sense) of the NS gene joined to a hepatitis delta virus ribozyme at its 3' end. This assay demonstrated that all deletion mutants of PB1 exhibited either background or greatly reduced polymerase activity irrespective of the ability to interact with PB2 and that all linker-insertion mutants except one at the extreme COOH end (L-746) of PB1 were also negative for viral polymerase activity. These results show that compared with complex formation of PB1 with PB2, the polymerase activity of PB1 was extremely sensitive to structural perturbation. 相似文献