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31.
32.
Glutamate 1-semialdehyde aminotransferase (glutamate 1-semialdehyde 2,1-aminomutase; EC 5.4.3.8; GSA-AT) catalyzes the transfer of the amino group on carbon 2 of glutamate 1-semialdehyde (GSA) to the neighboring carbon 1 to form delta-aminolevulinic acid (ALA). To gain insight into the mechanism of this enzyme, possible intermediates were tested with purified enzyme and the reaction sequence was followed spectroscopically. While 4,5-dioxovaleric acid (DOVA) was efficiently converted to ALA by the pyridoxamine 5'-phosphate (PMP) form of the enzyme, 4,5-diaminovaleric acid (DAVA) was a substrate for the pyridoxal 5'-phosphate (PLP) form of GSA-AT. Thus, both substances are reaction intermediates. The purified enzyme showed an absorption spectrum with a peak around 338 nm. Addition of PLP led to increased absorption at 338 nm and a new peak around 438 nm. Incubation of the purified enzyme with PMP resulted in an additional absorption peak at 350 nm. The reaction of the PLP and PMP form of the enzyme with GSA allowed the detection of a series of peaks which varied in their intensities in a time-dependent manner. The most drastic changes to the spectrum that were observed during the reaction sequence were at 495 and 540 nm. Some of the detected absorption bands during GSA-AT catalysis were previously described for several other aminotransferases, indicating the relationship of the mechanisms. The reaction of the PMP form of the enzyme with DOVA resulted in a similar spectrum as described above, while the spectrum for the conversion of DAVA by the PLP form of the enzyme indicated a different mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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Per Putkonen Charlotta Nilsson Kerstin Hild Reinhold Benthin Martin Cranage Anne Marie Aubertin Gunnel Biberfeld 《Journal of medical primatology》1993,22(2-3):100-103
Several groups have reported protection against experimental SIV infection in macaques immunized with a whole inactivated virus vaccine. The aim of the current study was to investigate whether five macaques vaccinated with whole inactivated SIV and previously shown to be protected against challenge with two divergent strains of SIV grown on human cells could resist challenge with a subsequent homologous SIV grown on macaque cells. We show here that this same vaccine did not protect when the challenge virus was grown on primary cells of monkey origin. 相似文献
35.
Synaptic Vesicle Traffic: Rush Hour in the Nerve Terminal 总被引:6,自引:0,他引:6
36.
The class I glutamine (Gln) tRNA synthetase interacts with the anticodon and acceptor stem of glutamine tRNA. RNA hairpin helices were designed to probe acceptor stem and anticodon stem-loop contacts. A seven-base pair RNA microhelix derived from the acceptor stem of tRNAGln was aminoacylated by Gln tRNA synthetase. Variants of the glutamine acceptor stem microhelix implicated the discriminator base as a major identity element for glutaminylation of the RNA helix. A second RNA microhelix representing the anticodon stem-loop competitively inhibited tRNAGln charging. However, the anticodon stem-loop microhelix did not enhance aminoacylation of the acceptor stem microhelix. Thus, transduction of the anticodon identity signal may require covalent continuity of the tRNA chain to trigger efficient aminoacylation. 相似文献
37.
Studies on the biology and function of human cytomegalovirus (HCMV) genes have been hampered by the limited number of viral mutants available for genetic analyses. We have developed a simple procedure to generate and enrich for HCMV recombinants. By inserting the bacterial neo gene, encoding neomycin/kanamycin phosphotransferase, into the large HCMV DNA genome using homologous recombination, selectable mutants of this complex herpesvirus were isolated for the first time. The synthesis of Neo from the viral genome was used to effectively enrich for recombinant viruses (re-viruses) in permissive culture cells grown in the presence of Geneticin (G418). A quick assay for Neo activity in infected cells, based on phosphorylation of kanamycin (Km), was used to easily identify viral recombinants in the process of screening and isolation. This procedure, not used previously to identify re-viruses, proved to be very useful for screening of large numbers of HCMV recombinants. Analysis of re-virus by Southern blotting revealed that the insertion of the marker gene had resulted in the expected deletion of the open reading frames, TRL 13/14 and UL 1–5, of HCMV. Re-virus was stable and showed no differences in growth kinetics as compared to wild-type (wt) virus. The insertion of a selectable marker gene into the HCMV genome and identification of viral recombinants by the Km phosphorylation assay, as presented here, provides the rationale for effective generation, enrichment and stable propagation of HCMV mutants. 相似文献
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39.
Reinhold Sulzmann 《Cell and tissue research》1956,44(1):87-100
Zusammenfassung Die Ansicht vonTheodor Boveri (1885), wonach sich dieSchwann-sche Scheide an denRanvierschen Schnürringen von der Außenseite des Markes auf dessen Innenseite umschlägt und so das Axolemm bildet, kann an Hand von formalinfixierten und gefärbten Präparaten (Serienlängsschnitten) bestätigt und durch entsprechende Mikroaufnahmen belegt werden.Der Begriff derSchwannschen Zelle (= Neurolemmzelle) ist demnach insofern jetzt weiter zu fassen, als diese einröhrenförmiges Gebilde darstellt, das die Markscheide in sich einschließt.Es wird deshalb erneut vorgeschlagen, dieSchwannsche Scheide fortan als äußeres Neurolemm und das Axolemm als inneres Neurolemm zu bezeichnen und auch die jeweils zugehörigen, bereits an anderer Stelle (R. Sulzmann 1955) beschriebenen Leisten dementsprechend zu benennen.Die Tatsache, daß die Schnürringe von Fasern nahezu gleicher Stärke in ein und demselben Faserbündel stets auf annähernd gleicher Höhe anzutreffen sind, wird lediglich als eineFolge gleicher Wachstumsgeschwindigkeit ohne besondere funktionelle Bedeutung aufgefaßt. Bezüglich der Funktion der Schnürringe wird vermutet, daß sie in erster Linie derErnährung der jeweils angrenzendenSchwannschen Segmente, insbesondere aber dem Stoffaustausch der betreffenden Achsenzylinderabschnitte dienen.Weiterhin wird angenommen, daß sich die Besonderheiten im Bau desRanvierschen Schnürrings, die zweifelsohne von großer praktischer Bedeutung sind, aus derEntwicklungsgeschichte der peripheren markhaltigen Nervenfaser ableiten lassen. 相似文献
40.
Holger Hackstein Gerhard Jahn Holger Kirchner Gregor Bein 《Histochemistry and cell biology》1996,106(2):229-234
Radioactive in situ hybridization techniques or enzymatic detection procedures of hapten-modified human cytomegalovirus (HCMV)
probes have been widely used for studying the infection of peripheral blood leukocytes with HCMV. This report describes significant
improvements in terms of signal resolution which can be obtained by applying a highly sensitive fluorescence in situ hybridization
(FISH) technique in conjunction with a large subgenomic HCMV DNA probe. Three cosmid clones spanning 119.1 kb of the HCMV
genome (230 kb) were used to construct the digoxigenin-11-dUTP-labeled probe which was found to be superior to a total HCMV
probe representing the entire genome. Crucial hybridization parameters were analyzed systematically in order to ensure optimal
resolution power and sensitivity. The protocol was successfully applied to HCMV-infected fibroblasts and peripheral blood
leukocytes of 12 transplant patients and unambiguously facilitated the precise intracellular localization of HCMV genomes
in infected cells. Because of its excellent resolution properties, accompanied by the virtual absence by any background staining,
we recommend the use of this protocol as a sensitive approach for further virological analyses of the interactions between
HCMV and peripheral blood leukocytes at the single-cell level.
Accepted: 16 February 1996 相似文献