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41.
Howard R. Morris Karen E. Batley Nigel G. L. Harding Richard A. Bjur John G. Dann Rodney W. King 《The Biochemical journal》1974,137(2):409-411
An elastase digest of a protein of unknown structure, dihydrofolate reductase, was studied by mass spectrometry. This soluble digest contained a large number of small peptides in different yields, within the ideal molecular-weight range (200-1200) for mixture-analysis mass spectrometry. Sequences of the major component peptides in the digest are reported. 相似文献
42.
43.
James M. Berger Paul J. Jackson Nigel J. Robinson Leah D. Lujan Emmanuel Delhaize 《Plant cell reports》1989,7(8):632-635
Suspension cultures of Datura innoxia cells were pulse-labeled with [35S]cysteine, then exposed to Cd to determine whether there is a direct precursor-product relationship amongst the different forms of the Cd-induced polypeptides, poly(-glutamylcysteinyl)glycines [(EC)nG, n=2 to 5]. Degradation of the polypeptides and possible regeneration of the [35S]-labeled glutathione and cysteine pools were also examined. After 2 h of exposure to [35S]cysteine, about 70% of the [35S]cysteine in the soluble fraction of the cell was incorporated into [35S]glutathione before exposure of the cells to Cd. One h after Cd exposure, most of the cellular [35S]glutathione was depleted and label was incorporated into (EC)nG. Analysis of [35S](EC)nG by reverse phase HPLC showed no direct precursor-product relationship between the synthesis of the shorter and longer chain forms. However, the rate of synthesis of the different polypeptides was linear for 32 h after Cd exposure. There was no evidence of degradation of [35S](EC)nG nor was it excreted into the medium within this period. From these results it is suggested that in the presence of Cd, a large pool of (EC)nG is unavailable for elongation to (EC)n+1G.Abbreviations (EC)nG
Poly(-glutamylcysteinyl)glycine
- HPLC
High pressure liquid chromatography
- CPM
Counts per minute 相似文献
44.
Nigel J. Robinson 《Journal of applied phycology》1989,1(1):5-18
Metallothioneins, MT's, are low-molecular-weight, cysteine-rich, polypeptides that complex ‘soft’ metal ions in thiol clusters.
They are structurally diverse. Some MT's are gene products, while others are secondary metabolites. Two of the three classes
of MT have been identified in algae. Eukaryotic algae possess the secondary metabolites referred to as class III MT. There
is no unequivocal evidence that MT genes occur in eukaryotic algae. However, the products of MT genes have been identified
in cyanobacteria. These genes and their metal regulatory elements remain to be isolated and characterized.
MT's have attracted interest from researchers involved in a wide range of disciplines including bioinorganic chemistry, biochemistry,
molecular biology, physiology, toxicology, environmental science and medicine. Although, the precise physiological roles of
these polypeptides remain undefined, a large number of functions have been speculated. These molecules chelate toxic trace
metals, such as Cd, thereby reducing the concentration of cytotoxic, free-metal ions. Furthermore, some MT's are believed
to be involved in zinc and copper homoeostasis. Future studies should reveal whether or not some of the diversity of MT structure
reflects a diversity of function. 相似文献
45.
Adelina A. Davies Stephen E. Moss Mark R. Crompton Tania A. Jones Nigel K. Spurr Denise Sheer Christine Kozak Michael J. Crumpton 《Human genetics》1989,82(3):234-238
Summary The gene encoding a tissue inhibitor of metalloproteinases, TIMP, has previously been shown to be X-linked in both the human and mouse genomes. We have used a series of somatic cell hybrids segregating translocation and deletion X chromosomes to map the TIMP gene on the human X chromosome. In combination with previous data, the gene can be assigned to Xp11.23Xp11.4. Genetic linkage analyses demonstrate that TIMP is linked to the more distal ornithine transcarbamylase (OTC) locus at a distance of about 22 centimorgans. The data are consistent with the conclusion that TIMP maps to a conserved synteny and linkage group on the proximal short arm of the human X chromosome and on the pericentric region of the mouse X chromosome, including loci for synapsin-1, a member of the raf oncogene family, OTC, and TIMP. 相似文献
46.
The small (116 amino acids) inner membrane protein MerT encoded by the transposon Tn501 has been overexpressed under the control of the bacteriophage T7 expression system. Random mutants of MerT were made and screened for loss of mercuric ion hypersensitivity. Several mutantmerT genes were selected and sequenced: Cys24Arg and Cys25Tyr mutations abolish mercury resistance, as do charge-substitution mutations in the first predicted transmembrane helix (Glyl4Arg, Glyl5Arg, Gly27Arg, Ala18Asp), and the termination mutations Trp66Ter and Cys82Ter. 相似文献
47.
Synopsis One perspective emphasizing the importance of stochastic processes in determining coral reef fish assemblages implies that there is little organization in species richness, abundance structure, and spatial distribution. We examine the degree to which this perspective is correct by analyzing distribution of fishes on a collection of patch reefs (Discovery Bay, Jamaica). We ask the question whether these patches accumulate species and individuals in a manner consistent with stochastic expectations. To address this question we use two conceptual models, each permitting a different insight. One assumes that fish are distributed stochastically on patches while the other assumes presence of restrictions on fish distribution due to habitat structure. For each conceptual model we use two types of benchmark: we compare observed patterns to those predicted by theoretical models, and we also compare observed patterns to those obtained from a random reallocation of fish individuals to patches. We found that the conceptual model assuming stochastic processes appeared to provide weaker explanation of patterns than the conceptual model that includes restrictions due to habitat structure. Further, and more importantly, we found that (i) the community is shaped by a mixture of stochastic and non-stochastic mechanisms, and (ii) the stochastic assembly processes decrease in importance for species restricted to fewer microhabitat types and sites. Our study therefore indicates that patches do accumulate individuals and species in a manner consistent with stochastic expectations, however, this applies primarily to the habitat generalists (unrestricted species). By the same token, increased habitat specialization by some species imposes constraints on the stochastic model such that it eventually fails. 相似文献
48.
49.
Summary Peptides labelled with the fluorophore cyanine 3 were used to study naturally expressed neuropeptide receptors by confocal
microscopy in continuous cell lines, primary cultures, and unfixed tissue. Swiss 3T3 fibroblasts bound cyanine 3-gastrin-releasing
peptide at 4°C, and internalized the peptide after 10 min at 37°C. Internalization was specific, since it was blocked by incubation
with unlabelled peptide. Primary cultures of myenteric neurons of the guinea pig incubated with cyanine 3-substance P at 4°C
had specific surface labelling. After 30 s at 37°C, the peptide was internalized into vesicles in both the soma and neurites.
Direct observation of live neurons showed movement of fluorescent vesicles to a perinuclear region after 30 min. Endocytosis
was associated with a loss of surface binding sites. Unfixed whole mounts of guinea pig and rat ileum were incubated with
cyanine 3-neurokinin A at 4°C. After 5 min at 37°C, Cy3-neurokinin A was specifically internalized in neurons and smooth muscle
cells. After 30 min, a perinuclear labelling occurred in some cells. Labelling in rat neurons was diminished by the NK3-R
antagonist SR142801. Thus, cyanine 3-neuropeptides are valuable tools to study expression and endocytosis of naturally expressed
receptors. 相似文献
50.
Nienow AW Langheinrich C Stevenson NC Emery AN Clayton TM Slater NK 《Cytotechnology》1996,22(1-3):87-94
Because of concern for cell damage, very low agitation energy inputs have been used in industrial animal cell bioreactors, typical values being two orders of magnitude less than those found in bacterial fermentations. Aeration rates are also very small. As a result, such bioreactors might be both poorly mixed and also unable to provide the higher oxygen up-take rates demanded by more intensive operation. This paper reports experimental studies both of K
L
a and of mixing (via pH measurements) in bioreactors up to 8 m3 at Wellcome and of scaled down models of such reactors at Birmingham. Alongside these physical measurements, sensitivity of certain cell lines to continuously controlled dO2 has been studied and the oxygen up-take rates measured in representative growth conditions. An analysis of characteristic times and mixing theory, together with other recent work showing that more vigorous agitation and aeration can be used especially in the presence of Pluronic F-68, indicates ways of improving their performance. pH gradients offer a special challenge. 相似文献