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41.
42.
An assay for ferredoxin-glutamate synthase is introduced thatuses an anion exchange resin to isolate the glutamate formedand subsequent determination with the ninhydrin procedure. Theenzyme was purified 200-fold from corn leaves by ammonium sulfatefractionation and chromatography on DEAE-cellulose, DEAE-Sephaceland ferredoxin- Sepharose. The purified enzyme had a specificactivity of 14 µmoles glutamate formed min1mg1protein. The enzyme has a molecular weight of 160,000. The pHoptimum for catalytic activity is 6.9. The isoelectric pointis at pH 4.2. The apparent Km values of the enzyme for L-glutamine,2-oxoglutarate and ferredoxin are 1,100, 240 and 1.7 µM.The enzyme has a high specificity toward these substrates witha stoichiometry between glutamate formation and glutamine consumption.Sulfhydryl reagents, bathophenanthroline, phthalein acids andazaserine produced strong inhibition of the enzyme activity.
1Permanent address: Department of Agricultural Chemistry, KyotoUniversity, Kyoto 606, Japan.
2To whom inquiries should be addressed. (Received July 7, 1979; ) 相似文献
43.
Toru Shimizu Masahiro Hatano Seiji Nagao Yoshinori Nozawa 《Biochemical and biophysical research communications》1982,106(4):1112-1118
43Ca NMR spectra of Ca2+-Tetrahymena calmodulin(Tet. CaM.) complexes have been observed under various conditions. Off-rate of Ca2+ from Tet. CaM. is estimated to be approx. 2.7 × 103 s?1 under a certain assumption. Relaxation rates of 43Ca NMR of Ca2+-Tet. CaM. are remarkably increased(by one order in magnitude) by adding trifluoperazine(TFP), a potent calmodulin antagonist. Relaxation parameters estimated suggest that Ca2+ mobility is reduced by the TFP binding. A stoichiometry of TFP is two moles per Tet. CaM. molecule. The relaxation rates of 43Ca NMR signals are increased by adding excessive Mg2+ to the Ca2+-Tet. CaM. solutions. The addition of Mg2+ to the Ca2+-Tet. CaM. complex decreases apparent pKa value of the complex as well. 相似文献
44.
45.
Kunshan Gao Yusho Aruga Kozi Asada Toshiaki Ishihara Toru Akano Masataka Kiyohara 《Journal of applied phycology》1991,3(4):355-362
Leafy thalli of the red algaPorphyra yezoensis Ueda, initiated from conchospores released from free-living conchocelis, were cultured using aeration with high CO2. It was found that the higher the CO2 concentration, the faster the growth of the thalli. Aeration with elevated CO2 lowered pH in dark, but raised pH remarkably in light with the thalli, because the photosynthetic conversion of HCO 3 ? to OH? and CO2 proceeded much faster than the dissociation of hydrated CO2 releasing H+. Photosynthesis of the alga was found to be enhanced in the seawater of elevated dissolved inorganic carbon (DIC, CO2 + HCO 3 ? + CO 3 ? ). It is concluded that the increased pH in the light resulted in the increase of DIC in the culture media, thus enhancing photosynthesis and growth. The relevance of the results to removal of atmospheric CO2 by marine algae is discussed. 相似文献
46.
The product of the malE—lacZ gene fusion was reported to compete with some proteins including outer membrane lipoprotein in the protein translocation across the Echerichia coli membrane. The fusion product also inhibited colicin E1 export. Furthermore, globomycin, which accumulated prolipoprotein in the membrane, inhibited the translocation of colicin E1 in the wild-type cells, but not in lipoprotein-negative mutant cells. Since colicin E1 contains the internal signal-like sequence [Proc. Natl. Acad. Sci. USA (1982) 79, 2827–2831], these results suggest that colicin E1 is exported by the aid of this sequence at a common site for maltose-binding protein and lipoprotein translocation. 相似文献
47.
Degradation of Xyloglucan by Wall-bound Enzymes from Soybean Tissue I. Occurrence of Xyloglucan-degrading Enzymes in Soybean Cell Wall 总被引:1,自引:0,他引:1
Koyama Toru; Hayashi Takahisa; Kato Yoji; Matsuda Kazuo 《Plant & cell physiology》1981,22(7):1191-1198
An enzyme preparation that catalyzes the degradation of xyloglucanwas obtained by extraction of the cell walls of soybean hypocotylswith a buffer containing 1.0 M NaCl. The enzyme preparationwas shown to catalyze two-step degradation of xyloglucan. Thepolysaccharide was first degraded into comparatively large fragments,which were then further degraded into monosaccharides. In orderto elucidate the mode of degradation of the xyloglucan duringcell growth, the activities of xyloglucandegrading enzymes ofsoybean-hypocotyl segments were assayed at different stagesof elongation. The total activities of the degrading enzymeswere lower in the elongating regions than in the non-elongatingregions. However, high levels of endo-ß-l,4-glucanasewere found in the elongating regions. These results suggestthat xyloglucan is hydrolyzed by endo-ß-1,4-glucanaseinto comparatively large fragments at the initial stage of growthand the resulting fragments are further degraded into monosaccharidesduring cell elongation. (Received May 20, 1981; Accepted August 8, 1981) 相似文献
48.
Hideaki Yamada Toru Nagasawa Haruyuki Ohkishi Bunsei Kawakami Yoshiki Tani 《Biochemical and biophysical research communications》1981,100(3):1104-1110
Synthesis of D-cysteine from 3-chloro-D-alanine and hydrogen sulfide is catalyzed by highly purified 3-chloro-D-alanine hydrogen chloride-lyase from . The synthetic reaction proceeds optimally at pH 8.5, as a function of enzyme concentration and incubation time. The enzymatically synthesized D-cysteine was isolated from the large scale reaction mixture and identified by physicochemical means. 相似文献
49.
Cytochrome c oxidase from rat liver was incubated with various proteinases of different specificities and the enzymic activity was measured after various incubation times. A loss of catalytic activity was found after digestion with proteinase K, aminopeptidase M and a mitochondrial proteinase from rat liver. In each case the decrease in enzymic activity was compared with the changes in intensities of the polypeptide pattern obtained after sodium dodecyl sulfate polyacrylamide gel electrophoresis. The susceptibilities of the subunit polypeptides of the soluble cytochrome c oxidase to proteinases were very different. Whereas subunit I was most susceptible, subunits V–VII were rather resistant to degradation. From the relative inaccessibility of subunits V–VII to proteinases it is likely that these polypeptides are buried in the interior of the enzyme complex. 相似文献
50.
Circular dichroism (CD) and absorption spectra of squid (Todarodes pacificus) rhodopsin, isorhodopsin and the intermediates were measured at low temperatures. Squid rhodopsin has positive CD bands at wavelengths corresponding the - and β-absorption bands at liquid nitrogen temperature (CD maxima: 485 nm at -band and 348 nm at β-band) as well as at room temperature (CD maxima: 474 nm at -band and 347 nm at β-band). The rotational strength of the -band has a molecular ellipticity about twice that of cattle rhodopsin. The CD spectrum of bathorhodopsin displays a negative peak at 532 nm, the rotational strength of which has an absolute value slightly larger than that of rhodopsin. The reversal in sign at -band of the CD spectrum may indicate that the isomerization of retinal chromophore from twisted 11-cis form to twisted 11-trans form has occurred in the process of conversion from rhodopsin to bathorhodopsin. Lumirhodopsin has a small negative CD band at 490 nm, the maximum of which lies at 25 nm shorter wavelengths than the absorption maximum (515 nm), and a large positive CD band near 290 nm, which is not observed in rhodopsin and the other intermediates. This band may be derived from a conformational change of the opsin. In the process of changing from lumirhodopsin to LM-rhodopsin, the CD bands at visible and near ultraviolet regions disappear. Both alkaline and acid metarhodopsins have no CD bands at visible and near ultraviolet regions. 相似文献