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11.
J. L. Marco L. A. Bataus F. F. Valência C. J. Ulhoa S. Astolfi-Filho C. R. Felix 《Applied microbiology and biotechnology》1996,44(6):746-752
ABacillus subtilis amylase gene was inserted into a plasmid which transferred toEscherichia coli. During cloning, a 3 region encoding 171 carboxyterminal amino acids was replaced by a nucleotide sequence that encoded 33 amino acid residues not present in the indigenous protein. The transformed cells produced substantial amylolytic activity. The active protein was purified to apparent homogeneity. Its molecular mass (48 kDa), as estimated in sodium dodecyl sulfate/polyacrylamide gel electrophoresis, was lower than the molecular mass values calculated from the derived amino acid sequences of theB. subtilis complete -amylase (57.7 kDa) and the truncated protein (54.1 kDa). This truncated enzyme form hydrolysed starch with aK
m of 3.845 mg/ml. Activity was optimal at pH 6.5 and 50°C, and the purified enzyme was stable at temperatures up to 50°C. While Hg2+, Fe3+ and Al3+ were effective in inhibiting the truncated enzyme Mn2+ and Co2+ considerably enhanced the activity. 相似文献
12.
A new gene coding for an antigen recognized by autologous cytolytic T lymphocytes on a human renal carcinoma 总被引:7,自引:0,他引:7
Béatrice Gaugler Nathalie Brouwenstijn Valérie Vantomme Jean-Pierre Szikora Corry W. Van der Spek Jean-Jacques Patard Thierry Boon Peter Schrier Benoît J. Van den Eynde 《Immunogenetics》1996,44(5):323-330
Previous reports have described antigens that are recognized on human melanoma cells by autologous cytolytic T lymphocytes
(CTL). The genes coding for a number of these antigens have been identified. Here we report the cloning of a gene that codes
for an antigen recognized by autologous CTL on a human renal carcinoma cell line. This antigen is presented byHLA-B7 and is encoded by a new gene that we have namedRAGE1. No expression ofRAGE1 was found in normal tissues other than retina. RAGE1 expression was found in only one of 57 renal cell carcinoma samples,
and also in some sarcomas, infiltrating bladder carcinomas, and melanomas. This represents the first identification of an
antigen recognized by autologous CTL on a renal tumor. 相似文献
13.
Ghislain Marc Frankard Valérie Vandenbossche Dirk Matthews Benjamin F. Jacobs Michel 《Plant molecular biology》1994,24(6):835-851
The gene encoding Arabidopsis thaliana aspartate kinase (ATP:L-aspartate 4-phosphotransferase, EC 2.7.2.4) was isolated from genomic DNA libraries using the carrot ak-hsdh gene as the hybridizing probe. Two genomic libraries from different A. thaliana races were screened independently with the ak probe and the hsdh probe. Nucleotide sequences of the A. thaliana overlapping clones were determined and encompassed 2 kb upstream of the coding region and 300 bp downstream. The corresponding cDNA was isolated from a cDNA library made from poly(A)+-mRNA extracted from cell suspension cultures. Sequence comparison between the Arabidopsis gene product and an AK-HSDH bifunctional enzyme from carrot and from the Escherichia coli thrA and metL genes shows 80%, 37.5% and 31.4% amino acid sequence identity, respectively. The A. thaliana ak-hsdh gene is proposed to be the plant thrA homologue coding for the AK isozyme feedback inhibited by threonine. The gene is present in A. thaliana in single copy and functional as evidenced by hybridization analyses.The apoprotein-coding region is interrupted by 15 introns ranging from 78 to 134 bp. An upstream chloroplast-targeting sequence with low sequence similarity with the carrot transit peptide was identified. A signal sequence is proposed starting from a functional ATG initiation codon to the first exon of the apoprotein. Two additional introns were identified: one in the 5 non-coding leader sequence and the other in the putative chloroplast targeting sequence. 5 sequence analysis revealed the presence of several possible promoter elements as well as conserved regulatory motifs. Among these, an Opaque2 and a yeast GCN4-like recognition element might be relevant for such a gene coding for an enzyme limiting the carbon-flux entry to the biosynthesis of several essential amino acids. 3 sequence analysis showed the occurrence of two polyadenylation signals upstream of the polyadenylation site.This work is the first report of the molecular cloning of a plant ak-hsdh genomic sequence. It describes a promoter element that may bring new insights to the regulation of the biosynthesis of the aspartate family of amino acids.Abbreviations AK
aspartate kinase
- HSDH
homoserine dehydrogenase
- ID
intermediate domain
- Tp
transit peptide 相似文献
14.
J. Del Val Pérez J. E. Fa J. Castroviejo F. J. Purroy 《Biodiversity and Conservation》1994,3(9):868-892
The status of 50 taxonomically unique bird taxa found in Bioko is described. A complete updated species list for birds in Bioko is also given. Notes on habitat occupied and altitudinal distribution of each species as well as information on their general abundance is also included. Species distributions may have been affected by habitat modifications brought about by humans but most taxonomically unique forms are found in the montane areas. Comparisons are made with the neighbouring Mount Cameroon area. 相似文献
15.
Martine Crasnier Valérie Dumay Antoine Danchin 《Molecular genetics and genomics : MGG》1994,243(4):409-416
In Escherichia coli, adenylate cyclase activity is regulated by phosphorylated EnzymeIIAGlc, a component of the phosphotransferase system for glucose transport. In strains deficient in EnzymeIIAGlc, CAMP levels are very low. Adenylate cyclase containing the D414N substitution produces a low level of cAMP and it has been proposed that D414 may be involved in the process leading to activation by EnzymeIIAGlc. In this work, spontaneous secondary mutants producing large amounts of cAMP in strains deficient in EnzymeIIAGlc were obtained. The secondary mutations were all deletions located in the cya gene around the D414N mutation, generating adenylate cyclases truncated at the carboxyl end. Among them, a 48 kDa protein (half the size of wild-type adenylate cyclase) was shown to produce ten times more cAMP than wild-type adenylate cyclase in strains deficient in EnzymeIIAGlc. In addition, this protein was not regulated in strains grown on glucose and diauxic growth was abolished. This allowed the definition of a catalytic domain that is not regulated by the phosphotransferase system and produces levels of cAMP similar to that of regulated wild-type adenylate cyclase in wild-type strains grown in the absence of glucose. Further analysis allowed the characterization of the COOH-terminal regulatory domain, which is proposed to be inhibitory to the activity of the catalytic domain. 相似文献
16.
Aurintricarboxylic Acid Protects Hippocampal Neurons from NMDA-and Ischemia-Induced Toxicity In Vivo 总被引:2,自引:1,他引:1
Jill M. Roberts-Lewis Val R. Marcy Yonghua Zhao Jeffry L. Vaught Robert Siman Michael E. Lewis 《Journal of neurochemistry》1993,61(1):378-381
Abstract: The polymeric dye aurintricarboxylic acid (ATA) has been shown to protect various cell types from apoptotic cell death, reportedly through inhibition of a calcium-dependent endonuclease activity. Recent studies have indicated that there may be some commonalities among apoptosis, programmed cell death, and certain other forms of neuronal death. To begin to explore the possibility of common biochemical mechanisms underlying ischemia-or excitotoxin-induced neuronal death and apoptosis in vivo, gerbils or rats subjected to transient global ischemia or NMDA microinjection, respectively, received a simultaneous intracerebral infusion of ATA or vehicle. As a biochemical marker of neuronal death, spectrin proteolysis, which is mediated by activation of calpain I, was measured in hippocampus after 24 h. ATA treatment resulted in a profound reduction of both NMDA-and ischemia-induced spectrin proteolysis, consistent with the possibility of some common mechanism in apoptosis and other forms of neuronal death in vivo. 相似文献
17.
John S. Beck Anne E. Kwitek Phillip H. Cogen Andrew K. Metzger Geoffrey M. Duyk Val C. Sheffield 《Human genetics》1993,91(1):25-30
p53 is a tumor suppressor gene located on 17p, a region of the human genome frequently deleted in tumors. Mutation of the p53 gene is an important step leading to development of many forms of human cancer. To simplify the analysis of tumors for p53 point mutations, we describe a GC-clamped denaturing gradient gel assay for detecting single-base substitutions within highly conserved regions of the p53 gene. This assay alows for efficient screening of tumors for single-base substitutions within the p53 gene and can be used to facilitate sequence analysis of p53 point mutations. 相似文献
18.
Valéria Marques Gabriel Riaño Miguel A. Carretero Iolanda Silva-Rocha Catarina Rato 《Acta zoologica》2023,104(3):419-433
Under temperature sex determination (TSD), sex is determined by temperature during embryonic development. Depending on ecological and physiological traits and plasticity, TSD species may face demographic collapse due to climate change. In this context, asymmetry in bilateral organisms can be used as a proxy for developmental instability and, therefore, deviations from optimal incubation conditions. Using Tarentola mauritanica gecko as a model, this study aimed first to confirm TSD, its pattern and pivotal temperature, and second to assess the local adaptation of TSD and variation of asymmetry patterns across four populations under different thermal regimes. Eggs were incubated at different temperatures, and hatchlings were sexed and measured. The number of lamellae was counted in adults and hatchlings. Results were compatible with a TSD pattern with males generated at low and females at high incubation temperatures. Estimated pivotal temperature coincided with the temperature producing lower embryonic mortality, evidencing selection towards balanced sex ratios. The temperature of oviposition was conservatively selected by gravid females. Asymmetry patterns found were likely related to nest temperature fluctuations. Overall, the rigidity of TSD may compromise reproductive success, and demographic stability in this species in case thermal nest choice becomes constrained by climate change. 相似文献
19.
Valérie Copié John A. Battles John M. Schwab Dennis A. Torchia 《Journal of biomolecular NMR》1996,7(4):335-340
Summary Nearly complete backbone 1H, 15N and 13C signal assignments are reported for -hydroxydecanoyl thiol ester dehydrase, a 39-kDa homodimer containing 342 amino acids. Although 15N relaxation data show that the protein has a rotational correlation time of 18 ns, assignments were derived from triple-resonance experiments recorded at 500 MHz and pH 6.8, without deuteration. The Chemical Shift Index, CSI, identified two long helices and numerous -strands in dehydrase. The CSI predictions are in close agreement with the secondary structure identified in the recently derived crystal structure, particularly when one takes account of the numerous bulges in the -strands. The assignment of dehydrase and a large deuterated protein [Yamazaki et al. (1994) J. Am. Chem. Soc., 116, 11655–11666] suggest that assignment of 40–60 kDa proteins is feasible. Hence, further progress in understanding the chemical shift/structure relationship could open the way to determine the structures of such large proteins.
Supplementary Material is available on request, comprising Table S1 listing the spectral parameters; Table S2 listing the assignments; Fig. S1 showing the 2D 1H–15N HSQC spectrum; Fig. S2 showing sequential NOEs, secondary shifts, H-exchange and 3JHN
data; and Fig. S3 showing plots of the H, C, CO and C Chemical Shift Indexes.To whom correspondence should be addressed. 相似文献
20.
Rachel Baltz Jean-Luc Evrard Val/'erie Bourdon Andr/'e Steinmetz 《Sexual plant reproduction》1996,9(5):264-268
The protein PLIM-1 (formerly SF3) from sunflower is expressed exclusively in mature, free pollen. It contains two LIM domains associated with an acidic C-terminus comprising six copies of the pentapeptide motif (A,T,S) (E,D) TQN. We have expressed the pollen protein as well as some of its mutant forms inEscherichia coli and have used the bacterially produced proteins to study interactions with nucleic acids. Our studies show that the protein binds DNA and RNA in vitro to form large complexes, while mutant polypeptides containing either a single LIM domain or a destabilized first or second LIM domain do not. Although these data suggest that the biological function of PLIM-1 involves interactions with nucleic acids, its role in pollen development remains unclear. 相似文献