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1.
1H nuclear magnetic resonance (1H NMR) experiments on Co(II)-substituted stellacyanin have been performed. Large paramagnetic hyperfine shifts are observed, the whole spectrum covering a range of 190 ppm. Experiments were mainly performed at 270 MHz from which temperature and pH* dependencies of the out-shifted resonances were reported, as well as determinations of the longitudinal (T1) and transverse (T2) relaxation times. These relaxation times are among other things, dependent on the individual proton-metal distance, and the aim of this work has been to determine these distances, by use of the Solomon-Bloembergen equations modified to include the so-called "Curie spin". The application of this method to a protein has not been reported earlier. Experiments were also performed at 100, 400, and 500 MHz in order to estimate the size of the Curie spin from the field dependence of the line widths. Furthermore, determination of the values for the rotational correlation time, tau r, and the effective magnetic moment, mu eff, was necessary for the present approach. With apostellacyanin, tau r was found to be (6.0 +/- 0.4) X 10-8 s. From the paramagnetic susceptibility of Co(II) stellacyanin, the value (4.53 +/- 0.03)beta was determined for mu eff. The proposed assignments of several paramagnetically out-shifted resonances. the proton-metal distances obtained, and the known peptide sequence of stellacyanin have allowed us to build a three-dimensional model of the metal site and its surrounding structure consistent with all the experimental data. It is revealed that both histidine ligands bind the metal with their 3-nitrogens. Also we find strong indications that a second sulfur atom is actually binding the metal, this being the long-sought-after fourth ligand. The model suggests that this sulfur belongs to Cys-59, which together with Cys-93 constitutes the disulfide bridge known to be present in the structure. A potential fifth ligand, an amide oxygen from Asn-47, is also found. 相似文献
2.
Payant V; Abukashawa S; Sasseville M; Benkel BF; Hickey DA; David J 《Molecular biology and evolution》1988,5(5):560-567
Nuclear DNA was extracted from each of the eight species comprising the
Drosophila melanogaster species subgroup. Southern hybridization of this
DNA by using a molecular probe specific for the alpha-amylase coding region
showed that the duplicated structure of the amylase locus, first found in
D. melanogaster, is conserved among all species of the melanogaster
subgroup. Evidence is also presented for the concerted evolution of the
duplicated genes within each species. In addition, it is shown that the
glucose repression of amylase gene expression, which has been extensively
studied in D. melanogaster, is not confined to this species but occurs in
all eight members of the species subgroup. Thus, both the duplicated gene
structure and the glucose repression of Drosophila amylase gene activity
are stable over extended periods of evolutionary time.
相似文献
3.
Biosynthesis of the neural cell adhesion molecule: characterization of polypeptide C 总被引:15,自引:4,他引:11
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O Nybroe M Albrechtsen J Dahlin D Linnemann J M Lyles C J M?ller E Bock 《The Journal of cell biology》1985,101(6):2310-2315
The biosynthesis of the neural cell adhesion molecule (N-CAM) was studied in primary cultures of rat cerebral glial cells, cerebellar granule neurons, and skeletal muscle cells. The three cell types produced different N-CAM polypeptide patterns. Glial cells synthesized a 135,000 Mr polypeptide B and a 115,000 Mr polypeptide C, whereas neurons expressed a 200,000 Mr polypeptide A as well as polypeptide B. Skeletal muscle cells produced polypeptide B. The polypeptides synthesized by the three cell types were immunochemically identical. The membrane association of polypeptide C was investigated with methods that distinguish peripheral and integral membrane proteins. Polypeptide C was found to be a peripheral membrane protein, whereas polypeptides A and B were integral membrane proteins with cytoplasmic domains of approximately 50,000 and approximately 25,000 Mr, respectively. The affinity of the membrane binding of polypeptide C increased during postnatal development. The posttranslational modifications of polypeptide C were investigated in glial cell cultures, and it was found to be N-linked glycosylated and sulfated. 相似文献
4.
The expanding use of computers in radiation therapy procedures, especially the rapidly increasing use of digital CT-information, necessitates the coordination of the different systems in order to facilitate their developments. In order to define necessary demands for tomorrow a Nordic cooperation was initiated 1981 by NORDFORSK (Nordic co-operative organization for applied research), and a group of physicians and physicists having their daily work in this field of medicine and physics was invited to produce a report on 'User requirements on CT-based computed dose planning systems on radiation therapy'. The work has been done within the frame of NORDFORSK's activities and has been independent of the existing commissions and associations in the radiology field, but it has taken into consideration recommendations that have been given by or are being produced by other organizations. This report is a short summary of the complete paper which will be published in Acta Radiologica. The aim of this short version is to get an early presentation of the 'requirement lists' (see Appendix) which we think are of immediate importance. 相似文献
5.
JOÃO BATISTA TAVARES DA SILVA ISAAC ROITMAN 《The Journal of eukaryotic microbiology》1990,37(6):521-523
ABSTRACT. Three strains of Phytomonas serpens two from tomatoes, Lycopersicon esculentum one from the insect Phtia picta (Hemiptera, Coreidae), were cultivated in a chemically defined medium developed from a defined medium for cultivating insect flagellates. Besides organic growth factors required by other insect trypanosomatids this flagellate requires, serine and inositol. Glutamine stimulates growth, and, surprisingly, does not require heme. 相似文献
6.
MARC J. FAZIO ALBA C. DA SILVA THOM K. ROSIERE G. BENJAMIN BOUCK 《The Journal of eukaryotic microbiology》1995,42(5):570-580
ABSTRACT. Proteins of the membrane skeleton of Euglena gracilis were extensively phosphorylated in vivo and in vitro after incubation with [32 P]-orthophosphate or γ-[32 P] ATP. Endogenous protein threonine/serine activity phosphorylated the major membrane skeletal proteins (articulins) and the putative integral membrane protein (IP39) anchor for articulins. The latter was also the major target for endogenous protein tyrosine kinase activity. A cytoplasmic domain of IP39 was specifically phosphorylated, and removal of this domain with papain eliminated the radiolabeled phosphoamino acids and eliminated or radically shifted the PI of the multiple isoforms of IP39. In gel kinase assays IP39 autophosphorylated and a 25 kDa protein which does not autophosphorylate was identified as a threonine/serine (casein) kinase. Plasma membranes from the membrane skeletal protein complex contained threonine/serine (casein) kinase activity, and cross-linking experiments suggested that IP39 was the likely source for this membrane activity. pH optima, cation requirements and heparin sensitivity of the detergent solubilized membrane activity were determined. Together these results suggest that protein kinases may be important modulators of protein assembly and function of the membrane skeleton of these protistan cells. 相似文献
7.
Borrelia burgdorferi is a spirochete pathogen transmitted among warm-
blooded hosts by ixodid ticks. Frequency-dependent selection for variant
outer-surface proteins might be expected to arise in this species, since
rare variants are more likely to avoid immune surveillance in previously
infected hosts. We sequenced the OspA and OspB genes of nine North American
strains and compared them with nine strains previously described. For each
gene, the mean number of synonymous substitutions per synonymous site and
the mean number of nonsynonymous substitutions per nonsynonymous site show
only a twofold excess of silent mutations. Synonymous rates vary widely
along the OspB protein. Some regions show a significant excess of silent
substitutions, while divergence in other regions is constrained by biased
base composition or selection. The presence, in antigenically important
regions of the protein, of significant variation among strains, as well as
evidence for recombination among strains, should be considered in attempts
to develop vaccines against this disease.
相似文献
8.
M P ROBINSON G BUTCHER R H CURTIS K G DA VIES K EVANS 《The Annals of applied biology》1993,123(2):337-347
Two monoclonal antibodies, which differentially recognise the two species of potato cyst nematodes (PCN), Globodera pallida and G. rostochiensis, are described. They have been shown to have potential for quantification of these two species, recognising proteins of the same molecular weight (34 kD) in both species. Further investigation showed these proteins to have isoelectric points at pH values of 5.7 in G. pallida and 5.9 in G. rostochiensis, in common with the proteins used by Fleming & Marks (1983) to differentiate the species of PCN. They are likely to be structurally very similar, with the same physiological function (and therefore similar concentrations) in the two species. In cross-reactivity tests with a wide range of soil nematode species, the antibodies reacted strongly only with species of the genus Globodera, and thereby confirmed their potential as the basis of a quantitative immunoassay likely to be useful in management of PCN populations. 相似文献
9.
10.