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81.
Manfred Braun Jong Min Kim Rolf D. Schmid 《Applied microbiology and biotechnology》1992,37(5):594-598
Summary The soil isolate Cellulomonas cellulans AM8 produces an extracellular l-amino acid oxidase (L-AAO) with broad substrate specificity. The strain produced up to 0.35 unit (U)/ml of the extracellular L-AAO in a simple medium containing glycerol and yeast extract. The enzyme was easily purified up to 30 U/mg protein using Phenyl-Sepharose fast flow. The purified enzyme migrated as single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) with a molecular mass of 55 kDa. On native PAGE the molecular mass was approx. 300 000 kDa, which may be due to aggregation. With the exception of glycine, proline, and threonine, all the amino acids normally constituting proteins were oxidized. The V
max values from 0.7 to 35.2 U/mg for aspartic acid and lysine, respectively, and the K
m values from 0.007 to 7.1 mm for cysteine and valine, respectively, were obtained at 25° C and pH 7.0 in oxygen-saturated solutions. The L-AAO had a pH optimum of 6.5–7.5. It was stable for several months at — 30° C and for some days at 35° C. Ferricyanide served as an electron acceptor with a V
max of 50 U/mg and K
m for 0.3 mm with phenylalanine as the substrate.
Correspondence to: R. D. Schmid 相似文献
82.
B. H. Min 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1980,183(4)
A method is described for measuring a trimethyl prostaglandin E2 analog, TM-PGE2, in human plasma. Trideuterated and monofluorinated analogs of TM-PGE2 are added to plasma as internal standard and carrier, respectively. The plasma is adjusted to pH 3.0 and is extracted with a mixture of benzene—dichloromethane (9:1). The residue, following removal of the extracting solvent, is reacted consecutively with pentafluorobenzyl bromide and bistrimethylsilyltrifluoroacetamide. The excess derivatizing reagents are removed by evaporation, and an aliquot of the reconstituted residue is analyzed by capillary column gas chromatography using methane as the carrier gas. A quadrupole mass spectrometer is set to monitor in the gas chromatographic effluent the (M − C7H2F5)− fragmention of TM-PGE2 (m/e 449) and trideuterated TM-PGE2 (m/e 452) generated by methane negative chemical ionization. Quantitation of unknowns is based on a comparison of the m/e 449 to m/e 452 ion ratio in each unknown to that obtained from the analysis of control plasma spiked with known amounts of TM-PGE2 and fixed amounts of internal standard and carrier. The sensitivity limit of the assay is approximately 100 pg ml−1, which is equivalent to 1 pg injected. The assay was used to measure the concentration of TM-PGE2 in the plasma of two subjects following a single 10 μg kg−1 oral dose of the drug. 相似文献
83.
Antibodies against purified ( from the rectal gland of Squalus acanthias, as well as against its catalytic subunit, inhibited ouabain binding by as much as 50%. However, antibodies against the glycoprotein subunit did not inhibit ouabain binding. These data suggest that binding of antibody against the catalytic subunit to the enzyme either covers the ouabain binding site or destroys its conformation, while binding of antibody against the glycoprotein has no such effect. 相似文献
84.
Galium procurrens is described as a new diploid relic species from Montenegro/N. Albania and SW. Bulgaria. It is related to the tetraploidG. laevigatum and other diploid and polyploid taxa of theG. sylvaticum-group inhabiting European deciduous forests. 相似文献
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89.
Lee Joo-Hyoung Park Jong-Ho Park Sun-Hye Kim Sun-Hong Kim Jee Yon Min Jeong-Ki Lee Gyun Min Kim Yeon-Gu 《Applied microbiology and biotechnology》2018,102(11):4729-4739
Applied Microbiology and Biotechnology - Despite the relatively low transfection efficiency and low specific foreign protein productivity (qp) of Chinese hamster ovary (CHO) cell-based transient... 相似文献
90.
Longhe Xu Felipe Matsunaga Jin Xi Min Li Jingyuan Ma 《Journal of biomolecular structure & dynamics》2013,31(11):1833-1840
We recently demonstrated that the anionic detergent sodium dodecyl sulfate (SDS) specifically interacts with the anesthetic binding site in horse spleen apoferritin, a soluble protein which models anesthetic binding sites in receptors. This raises the possibility of other detergents similarly interacting with and occluding such sites from anesthetics, thereby preventing the proper identification of novel anesthetic binding sites. n-Dodecyl β-D-maltoside (DDM) is a non-ionic detergent commonly used during protein-anesthetic studies because of its mild and non-denaturing properties. In this study, we demonstrate that SDS and DDM occupy anesthetic binding sites in the model proteins human serum albumin (HSA) and horse spleen apoferritin and thereby inhibit the binding of the general anesthetics propofol and isoflurane. DDM specifically interacts with HSA (Kd?=?40?μM) with a lower affinity than SDS (Kd?=?2?μM). DDM exerts all these effects while not perturbing the native structures of either model protein. Computational calculations corroborated the experimental results by demonstrating that the binding sites for DDM and both anesthetics on the model proteins overlapped. Collectively, our results indicate that DDM and SDS specifically interact with anesthetic binding sites and may thus prevent the identification of novel anesthetic sites. Special precaution should be taken when undertaking and interpreting results from protein-anesthetic investigations utilizing detergents like SDS and DDM. 相似文献