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21.
Sonicated DNA has been covalently attached to Sepharose 4B by a carbodiimide method [Rickwood, P. (1972) Biochim. Biophys. Acta269, 47–50] which minimizes modification of the DNA and matrix. Columns of this material have been used to study the interaction between cationic polypeptides and DNA. When poly(l-lysine) is bound to such columns at low ionic strength and then eluted with a linear salt gradient the polypeptide elutes over a broad range of salt concentration, presumably reflecting different strengths of interaction with various sites on the DNA. The broadness of the elution profile cannot be attributed to heterogeneity in the poly(l-lysine) sample but rather to the content of various DNA sites. Studies with other polypeptides, poly(l-Lys79, l-Leu21) and poly-(l-Lys-l-Ala-Gly), as well as studies at different temperatures, have helped to clarify the possible roles of conformational mobility, polypeptide hydrophobicity, and the presence of contiguous lysines in determining the strength of interaction of polypeptides and proteins with DNA sites. 相似文献
22.
Hordur Kristjansson Cyril Ponnamperuma 《Origins of life and evolution of the biosphere》1980,10(2):185-192
The enzyme malate dehydrogenase (EC 1.1.1.37) from an extreme thermophileB. Caldolyticus was purified to about 91% homogeneity. The molar mass of the enzyme was determined as 73 000 daltons and it is composed of two subunits, each with a molar mass of 37 000. Initial velocity studies with oxaloacetic acid and NADH as substrates at pH 8.1, over a range of temperatures, indicate that the enzyme operates via a sequential type mechanism. Van't Hoff plots of the kinetic parameters displayed sharp changes in slope at characteristic temperatures, whereas the Arrhenius plot exhibited no such breaks over the temperature interval investigated. The enzyme was found to be stable at 41°C and lower temperatures. At 51°C and 59°C an almost immediate 20% reduction in activity was obtained, but no further inactivation occurred during the 60 min of incubation. At 59°C the enzyme lost 50% of its initial activity in about 38 s. High concentration of NADH was observed to greatly stabilize the enzyme at that temperature.It is suggested that the slope changes in the Van't Hoff plots and the stability profies at 51°C and 59°C are representative of a temperature induced conformational change in the enzyme.Proceedings of the Fourth College Park Colloquium on Chemical Evolution:Limits of Life, University of Maryland, College Park, 18–20 October 1978. 相似文献
23.
We report the first partial purification of nitrous oxide reductase, a unique and labile enzyme of denitrifying bacteria. The procedure, which required anaerobic conditions throughout, resulted in a 60-fold purification relative to crude lysate in the case ofParococcus denitrificans. The molecular weight was estimated by gel exclusion chromatography to be about 85,000. The partially purified enzyme is inactivated rapidly by O2, dithionite, and mercaptoethanol and is reversibly inhibited by moderate concentrations of common salts. Up to 80% of the original activity can be reconstituted following O2 inactivation by incubating the enzyme with reduced benzyl viologen for 2 to 3 h. TheV
max pH profile shows a broad maximum at pH 8. The enzyme is irreversibly retained by common anion exchangers in the range pH 7 to 8 but can be eluted in acceptable yield as one of the last components from an imidazole-based anion exchange material by means of a pH gradient. This behavior implies that nitrous oxide reductase is very acidic. Among the several peptides observed by sodium dodecyl sulfate slab electrophoresis, only two, with apparent molecular weights of 58,000 and 25,000, correlated closely with the activity of fractions eluted from the imidazole column. These two peptides together comprised about 30% of the total protein in the fractions with highest specific activity. 相似文献
24.
Dana A. Procsal Helen L. Henry Ernest J. Friedlander Anthony W. Norman 《Archives of biochemistry and biophysics》1977,179(1):229-234
The biological activity of 1α,24R,25-trihydroxyvitamin D3 [1α,24R,25(OH)3D3] was elevated in comparison to the hormonally active form of vitamin D3, 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3], in the rachitic chick in terms of its ability to (a) stimulate intestinal calcium absorption, (b) mobilize bone calcium, (c) induce intestinal calcium binding protein, (d) modulate the level of enzyme activity of the renal 25-OH-D3-1-hydroxylase system, and (e) interact with the intestinal cystosol-chromatin receptor system for the 1α,25(OH)2D3 receptor system. In each of these assays, the relative ratio of activity of 1α,24R,25(OH)3D3 to 1α,25(OH)2D3was (a) 25–50, (b) ca. 20, (c) 10, (d) 50, and (e) 36%, respectively. 相似文献
25.
Morphine and Cocaine Exert Common Chronic Actions on Tyrosine Hydroxylase in Dopaminergic Brain Reward Regions 总被引:9,自引:0,他引:9
We studied levels of tyrosine hydroxylase immunoreactivity and phosphorylation state in the ventral tegmental area (VTA) and nucleus accumbens (NAc) in an effort to understand better the mechanisms by which these brain reward regions are influenced by opiates and cocaine. In the VTA, chronic, but not acute, administration of either morphine or cocaine increased levels of tyrosine hydroxylase immunoreactivity by 30-40%, with no change observed in the relative phosphorylation state of the enzyme. In the NAc, chronic, but not acute, morphine and cocaine treatments decreased the phosphorylation state of tyrosine hydroxylase, without a change in its total amount. In contrast, morphine and cocaine did not regulate tyrosine hydroxylase in the substantia nigra or caudate/putamen, brain regions generally not implicated in drug reward. Morphine and cocaine regulation of tyrosine hydroxylase could represent part of a common biochemical basis of morphine and cocaine addiction and craving. 相似文献
26.
Dana Lepofsky 《Economic botany》1992,46(2):192-211
The Mussau Islanders of the Bismarck Archipelago in northeastern Papua New Guinea cultivate 26 indigenous tree species both for food and non-food uses in tree gardens surrounding the villages. Several trees which volunteer in these arboriculture zones are also tended and encouraged. I present ethnobotanical information on methods of cultivation, how and when the trees are harvested, preparation and use. The tree crop zone has a random spatial distribution overall, but some species within the zone are aggregated spatially. This clumping is related to the restricted habitat requirements of certain species, as well as individual species ’ reproductive traits. Coconut is the most frequent species in the arboriculture zone and is randomly distributed. Vertical stratification is attributable to the presence of a distinct herb layer and a subcanopy stratum of maturePandanus spp. The stratification of some species pairs results from the deliberate planting of subcanopy trees next to canopy trees. The introduction of exotic garden species and the increasing availability of western packaged foods is changing the composition of traditional Mussau Islands tree gardens. 相似文献
27.
Annual egg production was determined for Artemia monica in Mono Lake, California, from 1983 to 1987. Annual oviparous (overwintering cyst) production was 3 and 7 million cysts m–2 yr–1 in 1986 and 1987, respectively, as measured by in situ sediment traps. Cyst production for the entire five year period was calculated using Artemia census data and inter-brood duration derived from mixolimnetic temperature. These estimates ranged from 2 to 5 million cysts m–2 yr–1. This method underestimated annual production by 30%, when compared to estimates using sediment traps. Cyst production was similar during 1983–1986 and showed a significant increase in 1987, which was due primarily to a larger reproductive population later in the year. Recruitment into the adult populations of the following spring ranged between 1.4 to 3.2%. Overall abundance of this generation reflected the patterns in annual cyst production. Compensatory effects must operate on the second generation of each year, since summer populations were similar in all years despite differences in cyst production. 相似文献
28.
R. Dana Ono 《Zoomorphology》1982,99(2):131-144
Summary Modified branchiostegal rays 1 through 3 support the proximal end of the paired hyoid barbels in the beardfish (Beryciformes: Polymixiidae). The polymixiid barbel is unusual in that it has an unique intrinsic muscular system. Using silver impregnation and electron microscopic techniques, unencapsulated, free nerve endings were located within the tendon of the third modified branchiostegal ray. Branchiostegal rays 1 and 2 do not have any free nerve endings associated with their tendons, however. It is suggested that the free nerve endings are proprioceptors acting as stretch-sensitive mechanoreceptors, and that branchiostegal ray 3 acts as part of a sensory apparatus for monitoring the positional state of the barbel. Branchiostegal rays 1 and 2 merely provide support for the barbel.Abbreviations used in Figures
BA
barbel
-
br r 3
branchiostegal ray 3
-
IM
intermandibularis
-
IOP
interoperculum
-
LIM
interoperculomandibular ligament
-
MD
mandible
-
MX
maxilla
-
OP
operculum
-
PM
premaxilla
-
POP
preoperculum
-
SOP
suboperculum 相似文献
29.
Transposition of the kanamycin-resistance transposon Tn903 总被引:6,自引:0,他引:6
Ry Young Dana Smith Grillo Ralph Isberg Jeffrey Way Michael Syvanen 《Molecular & general genetics : MGG》1980,178(3):681-689
Summary The insertion of the kanamycin-resistance transposon, Tn903, into the Escherichia coli chromosome was studied. Tn903 is similar in structure to the well known transposons Tn5 and Tn10 in that it has a unique central sequence flanked by inverted repeat sequences extending more than a thousand base pairs. However, the central region of Tn903 has enough single-frame coding capacity only for the drug modifying enzyme, whereas Tn5 and Tn10 carry multigenic unique sequences. In this paper we demonstrate that two different classes of insertion event occur: (1) the first class is a complex event in which all or part of the genome of the bacteriophage lambda vector is co-inserted near the purE locus on the E. coli chromosome (11.7 min); (2) the second class appears to be a simple transposition event in which the transposon alone is inserted at relatively nonspecific sites in the chromosome, as has been described for Tn5 and Tn10. Furthermore both classes show dependency on homology-requiring recombination systems. We suggest that Tn903 transposes infrequently because it must utilize a recA-controlled host function, whereas Tn5 and Tn10 are recA-independent and encode similar but more active functions on the transposon DNA. 相似文献
30.
Antigens induce sensitized lymphocytes to undergo mitosis and to secrete soluble products, termed lymphokines, which modulate the immune response. Plant lectins are known to act as polyclonal lymphocyte mitogens and, in some cases, stimulate lymphocytes to produce lymphokines. In an effort to explore the relationship of specific cell surface glycoconjugates to the induction of mitosis and the production of lymphokine activities we have examined the ability of the mitogenic lectins, concanavalin A and Wistaria floribunda mitogen, and the nonmitogenic hemagglutinin from Wistaria floribunda seeds to stimulate the production of macrophage migration inhibition factor (MIF), macrophage chemotactic factor (CF), and lymphotoxin (LT). Concanavalin A causes lymphocytes to produce MIF and LT but no detectable CF activities. W. floribunda mitogen induces lymphocytes to produce soluble substances which exhibit all three lymphokine activities. The nonmitogenic W. floribunda agglutinin causes lymphocytes to produce MIF and CF but no observable LT activity. Within the sensitivity of the assays employed, the results indicate that mitogenesis is not a corequisite of the expression of either macrophage migration inhibition factor or lymphocyte-derived chemotactic factor but it may be associated with the induction of lymphotoxin. It is also apparent that the expression of each lymphokine activity is independent of the expression of the other lymphokines studied. 相似文献