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1.
Binding of prothrombin, prethrombin 1, prethrombin 2 and thrombin to fibrinogen-Sepharose was studied. Thrombin and prethrombin 2 bound to fibrinogen-Sepharose, while prethrombin 1 and prothrombin did not. Bound thrombin and prethrombin 2 were recovered from the column by eluting with 0.1 M NaCl/0.05 M Tris-HCl buffer (pH 7.4). The affinity of thrombin and prethrombin 2 to fibrinogen-Sepharose depended on ionic strength and reached a maximum at 50 mm concentration. Prethrombin 2 interacts with fibrinogen as well as thrombin; and prothrombin fragment 1.2 is not important in the formation of this complex. Thus, prethrombin 2, which is a precursor of thrombin without measurable enzymatic activity and which lacks the single cleavage at Arg-322-Ile-323 present in thrombin, has the same or very similar structural conformation as thrombin and has the same macromolecular substrate recognition site. These results confirm the earlier results that active center is not necessary in fibrinogen-thrombin interaction. 相似文献
2.
J M Kaminski D Smith D S Reid W Kennedy R S Jeyendran L J Zaneveld 《Biology of reproduction》1987,36(5):1170-1176
Certain aryl 4-guanidinobenzoates (AGs; inhibitors of proteinases, including the sperm enzyme acrosin) have been shown to be more potent vaginal contraceptives in rabbits and less toxic than nonoxynol-9, the active ingredient of most marketed vaginal contraceptive formulations. To determine if these AGs can contact sperm and inhibit acrosin when mixed with the entire human ejaculate for a short period of time (roughly imitating clinical conditions), the inhibitors were added to semen at various concentrations for 2 min, after which the seminal plasma and unbound inhibitor were removed from the sperm by Ficoll centrifugation. Subsequently, the total arginine amidolytic activity of the spermatozoa was determined spectrophotometrically after a combined treatment that resulted in extraction, proacrosin activation, and reaction with substrate. Dose-response curves were prepared. All AGs studied were effective inhibitors of the amidolytic activity under these conditions, with ED50 values (the dose levels at which half of the acrosin associated with 10(6) sperm is inhibited) ranging from 10(-5) to 10(-7) M. To determine the effect on the proteolytic activity of individual spermatozoa, the experiment was repeated with 4'-acetamidophenyl 4-guanidinobenzoate (AGB), and the protease released from the sperm was measured by the gelatin-plate assay. The inhibition results were similar to those obtained by extraction of the spermatozoa and measurement of amidolytic activity. Thus, when mixed with the human ejaculate, AGs interact rapidly with spermatozoa to inhibit both their arginine amidolytic and proteolytic activity (probably due primarily or only to inhibition of acrosin) and remain bound even after removal of the seminal plasma. These data encourage further study of the compounds for contraceptive purposes. 相似文献
3.
G P Akulov Iu L Kaminski? A D Shestakov V G Kaiumov L F Chernysheva 《Bioorganicheskaia khimiia》1988,14(1):37-42
Chemical and enzymatic syntheses of [5'-3H]adenosine, [5'-3H]guanosine, and [5'-3H]uridine have been developed. The reduction of beta-D-ribo-pentadialdo-1,4-furanosyl derivatives of corresponding bases is used in the chemical synthesis. The maximum molar activity of the labelled products was 220 TBk/mol in reactions with [3H]NaBH4 and 370-740 TBk/mol in reactions with gaseous tritium. The enzymatic synthesis was performed by the rebosylation of heterocyclic bases with nucleoside phosphorylase and [5'-3H]uridine as a ribosyl donor. Nucleoside phosphorylase is proposed to be used in the immobilized form to avoid the decrease of molar activity. Nucleosides labelled with tritium both in ribosyl and heterocyclic moieties were synthesised enzymatically. 相似文献
4.
The expression of a lectin gene in pea (Pisum sativum L.) roots has been investigated using the copy DNA of a pea seed lectin as a probe. An mRNA which has the same size as the seed mRNA but which is about 4000 times less abundant has been detected in 21-d-old roots. The probe detected lectin expression as early as 4 d after sowing, with the highest level being reached at 10 d, i.e. just before nodulation. In later stages (16-d- and 21-d-old roots), expression was substantially decreased. The correlation between infection by Rhizobium leguminosarum and lectin expression in pea roots has been investigated by comparing root lectin mRNA levels in inoculated plants and in plants grown under conditions preventing nodulation. Neither growth in a nitrate concentration which inhibited nodulation nor growth in the absence of Rhizobium appreciably affected lectin expression in roots.Abbreviation cDNA
copy DNA
- poly(A)+RNA
polyadenylated RNA 相似文献
5.
Fabrizio Arigoni P. Alexandre Kaminski Hauke Hennecke Claudine Elmerich 《Molecular & general genetics : MGG》1991,225(3):514-520
Summary The nucleotide sequence of a 4.1 kb DNA fragment containing the fixABC region of Azorhizobium caulinodans was established. The three gene products were very similar to the corresponding polypeptides of Rhizobium meliloti. The C-terminal domains of both fixB products displayed a high degree of similarity with the -subunits of rat and human electron transfer flavoproteins, suggesting a role for the FixB protein in a redox reaction. Two open reading frames (ORF) were found downstream of fixC. The first ORF was identified as fixX on the basis of sequence homology with fixX from several Rhizobium and Bradyrhizobium strains. The second ORF potentially encoded a 69 amino acid product and was found to be homologous to a DNA region in the Rhodobacter capsulatus nif cluster I. Insertion mutagenesis of the A. caulinodans fixX gene conferred a Nif– phenotype to bacteria grown in the free-living state and a Fix– phenotype in symbiotic association with the host plant Sesbania rostrata. A crude extract from the fixX mutant had no nitrogenase activity. Furthermore, data presented in this paper also indicate that the previously identified nifO gene located upstream of fixA was probably a homologue of the nifW gene of Klebsiella pneumoniae and Azotobacter vinelandii. 相似文献
6.
Synergistic antitumor effect of interferon and anti-idiotype monoclonal antibody in murine lymphoma 总被引:3,自引:0,他引:3
T Y Basham M S Kaminski K Kitamura R Levy T C Merigan 《Journal of immunology (Baltimore, Md. : 1950)》1986,137(9):3019-3024
Both IFN-alpha and anti-idiotype monoclonal antibody therapy have significant antitumor activity in vivo in a murine B cell lymphoma model. Combination therapy with syngeneic anti-idiotype antibody of the IgG2a or IgG2b isotype (a single i.p. injection of 100 micrograms) and recombinant human hybrid interferon-alpha A/D (10(4) to 10(6) U three times weekly for 3 wk) synergistically increased median survival time in mice challenged with a lethal dose of tumor cells compared with the sum of the median survival times of the two individual treatments. IFN-alpha has direct antiproliferative activity against 38C13 in vitro and enhances in vitro macrophage anti-idiotype antibody-specific cytolysis for IgG2a, IgG2b, and IgG1 isotypes. 相似文献
7.
Charles Kaminski 《Planta》1971,99(1):63-72
Summary The specific peroxidase (1.11.1.7) and phenoloxidase (1.10.3.1) activities are quantitatively measured during the life of Coleus, from germination until flowering. In most organs investigated, the peroxidase activity increases rapidly with growth while the phenoloxidase activity remains low. The latter activity is higher in root apices than in more differentiated regions of roots. From the results obtained it may be concluded that the phenoloxidase activity accompanies cellular proliferation. It is suggested that the peroxidase activity plays an indirect role in root initiation through its role in cellular differentiation.
Extrait d'une thèse de doctorat soutenue à la Faculté des Sciences de l'Université de Liège. 相似文献
Extrait d'une thèse de doctorat soutenue à la Faculté des Sciences de l'Université de Liège. 相似文献
8.
9.
Maike Petersen Elisabeth Häusler Juliane Meinhard Barbara Karwatzki Claudia Gertlowski 《Plant Cell, Tissue and Organ Culture》1994,38(2-3):171-179
Suspension cultures of Coleus blumei accumulate very high amounts of rosmarinic acid, an ester of caffeic acid and 3,4-dihydroxyphenyllactate, in medium with elevated sucrose concentrations. Since the synthesis of this high level of rosmarinic acid occurs in only five days of the culture period, the activities of the enzymes involved in the biosynthesis are very high. Therefore all the enzymes necessary for the formation of rosmarinic acid from the precursors phenylalanine and tyrosine could be isolated from cell cultures of Coleus blumei: phenylalanine ammonia-lyase, cinnamic acid 4-hydroxylase, hydroxycinnamoyl:CoA ligase, tyrosine aminotransferase, hydroxyphenylpyruvate reductase, rosmarinic acid synthase and two microsomal 3- and 3-hydroxylases. The main characteristics of these enzymes of the proposed biosynthetic pathway of rosmarinic acid will be described.Abbreviations DHPL
3,4-dihydroxyphenyllactate
- DHPP
3,4-dihydroxyphenylpyruvate
- pHPL
4-hydroxyphenyllactate
- pHPP
4-hydroxyphenylpyruvate
- RA
rosmarinic acid 相似文献
10.
The discovery of many noncanonical peptides detectable with sensitive mass spectrometry inside, outside, and on cells shepherded the development of novel methods for their identification, often not supported by a systematic benchmarking with other methods. We here propose iBench, a bioinformatic tool that can construct ground truth proteomics datasets and cognate databases, thereby generating a training court wherein methods, search engines, and proteomics strategies can be tested, and their performances estimated by the same tool. iBench can be coupled to the main database search engines, allows the selection of customized features of mass spectrometry spectra and peptides, provides standard benchmarking outputs, and is open source. The proof-of-concept application to tryptic proteome digestions, immunopeptidomes, and synthetic peptide libraries dissected the impact that noncanonical peptides could have on the identification of canonical peptides by Mascot search with rescoring via Percolator (Mascot+Percolator). 相似文献