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991.
Assembly and interactions of cotJ-encoded proteins, constituents of the inner layers of the Bacillus subtilis spore coat 总被引:2,自引:0,他引:2
Richard W. Seyler Jr Adriano O. Henriques Amanda J. Ozin & Charles P. Moran Jr 《Molecular microbiology》1997,25(5):955-966
During Bacillus subtilis endospore formation, a complex protein coat is assembled around the maturing spore. The coat is made up of more than two dozen proteins that form an outer layer, which provides chemical resistance, and an inner layer, which may play a role in the activation of germination. A third, amorphous layer of the coat occupies the space between the inner coat and the cortex, and is referred to as the undercoat. Although several coat proteins have been characterized, little is known about their interactions during assembly of the coat. We show here that at least two open reading frames of the cotJ operon ( cotJA and cotJC ) encode spore coat proteins. We suggest that CotJC is a component of the undercoat, since we found that its assembly onto the forespore is not prevented by mutations that block both inner and outer coat assembly, and because CotJC is more accessible to antibody staining in spores lacking both of these coat layers. Assembly of CotJC into the coat is dependent upon expression of cotJA . Conversely, CotJA is not detected in the coats of a cotJC insertional mutant. Co-immunoprecipitation was used to demonstrate the formation of complexes containing CotJA and CotJC 6 h after the onset of sporulation. Experiments with the yeast two-hybrid system indicate that CotJC may interact with itself and with CotJA. We suggest that interaction of CotJA with CotJC is required for the assembly of both CotJA and CotJC into the spore coat. 相似文献
992.
993.
Cariillo M.Belen; Milner Caroline M.; Ball Simon T.; Snoek Margriet; Campbell R.Duncan 《Glycobiology》1997,7(7):975-986
The Neu1 locus, in the S region of the murine histocompatibility-2complex, regulates the sialic acid content of several liverlysosomal enzymes. Three alleles, Neu1a, Neu1b, and Neu1c, havebeen described on the basis of differential sialylation of theenzyme liver acid phosphatase. The Neu1a allele occurs in asmall number of mouse strains, e.g., SM/J and is associatedwith sialidase deficiency. We recently described G9, a sialidasegene in the human major histocompatibility complex (Milner etal. (1997) J. Biol. Chem., 272, 45494558), and we nowreport the characterization of the equivalent gene in mouse.The protein product of the murine G9 gene is 409 amino acidsin length and is 83% identical to its human orthologue. Expressionof the murine G9 protein in insect cells has confirmed thatit is a sialidase, with optimal activity at pH 5. To elucidatethe basis of sialidase deficiency in mouse strains carryingthe Neu1a allele, we have sequenced the G9 coding regions frommice carrying the three Neu1 alleles and hence defined the aminoacid sequence characteristic of each allotype. Of particularinterest is a Leu-209 to Ile mutation that is unique to theNeu1a allotype and is associated with reductions in sialidaseactivity of 68% and 88% compared to the Neu1b and Neu1c allotypes,respectively, when these three protein variants are expressedin insect cells. Additional factors, such as differential expression,may also influence the activities of the Nen1 allotypes in vivo.We have observed that the level of G9 mRNA is substantiallyreduced in mice carrying the Neu1a allele compared to the Neu1b(8595% reduction) and Neu1c (70% reduction) alleles. H2 complex MHC Neu1 sialidase 相似文献
994.
Agrobacterium tumefaciens-mediated barley transformation 总被引:26,自引:2,他引:24
Sonia Tingay David McElroy Roger Kalla Sarah Fieg Mingbo Wang Sarah Thornton Richard Brettell 《The Plant journal : for cell and molecular biology》1997,11(6):1369-1376
Genetically transformed barley was produced by eco-cultivating immature embryo explants with Agrobacterium tumefaciens carrying a binary vector coding for chimaeric bacterial genes, bar and gus , and selecting for bialaphos-resistant cultures from which plants were regenerated. Integration of both genes was confirmed by gel blot hybridization analysis of DNA from the transformed plants and their progenies. From 1282 embryos, plants were recovered for 54 independently transformed lines, giving a transformation efficiency of 4.2%. Transgene numbers in the different lines ranged from single copy insertion to at least ten copies. Sixteen out of 18 plants grown to maturity were fully fertile. Both marker genes, bar and gus , were expressed and co-segregated in the T1 progeny plants. In the majority of cases, the genes showed Mendelian segregation predicted for transgene insertion at a single locus. In one family with multiple transgene insertions, molecular analysis of T1 and T2 plants suggested that the T-DNA had inserted at two unlinked loci. 相似文献
995.
Induction of alternative oxidase synthesis by herbicides inhibiting branched-chain amino acid synthesis 总被引:4,自引:0,他引:4
Serge Aubert Richard Bligny David A. Day James Whelan Roland Douce 《The Plant journal : for cell and molecular biology》1997,11(4):649-657
Sycamore suspension cells ( Acer pseudoplatanus L.) were incubated in the presence of sulfonylurea and imidazolinone herbicides. These inhibitors of acetolactate synthase (ALS), a key enzyme of branched-chain amino acid synthesis, triggered a dramatic induction of the alternative oxidase (AOX). AOX activity increased in treated cells, eventually exceeding cytochrome (cyt) pathway activity. This induction of AOX activity was correlated with the accumulation of a 35 kDa AOX protein in isolated mitochondria, detected by Western blotting with a monoclonal antibody against Sauromatum guttatum AOX. It was preceded by the accumulation of putative 1.6 kb AOX mRNA, detected using an Aox cDNA probe from soybean. The metabolic perturbations induced by the herbicides rather than the herbicide molecules themselves were responsible for this induction of AOX. However, α-oxobutyrate (one of the substrates of ALS) and its transamination product, α-aminobutyrate, which accumulated after herbicide treatment, were not involved. The inhibition of branched-chain amino acid synthesis was probably somehow responsible for the AOX induction since: (i) a mixture of those amino acids (leucine, isoleucine, valine) prevented AOX induction by ALS inhibitors; (ii) the herbicide Hoe 704, a potent inhibitor of acetolactate reducto-isomerase (the enzyme following ALS in the branched-chain amino acid pathway), also triggered AOX induction. 相似文献
996.
Richard Cooke Monique Raynal Michele Laudié Michel Delseny 《The Plant journal : for cell and molecular biology》1997,11(5):1127-1140
Partial cDNA sequencing to obtain expressed sequence tags (ESTs) has led to the identification of tags to about 8000 of the estimated 20 000 genes in Arabidopsis thaliana . This figure represents four to five times the number of complete coding sequences from this organism available in international databases. In contrast to mammals, many proteins are encoded by multigene families in A. thaliana . Using ribosomal protein gene families as an example, it is possible to construct relatively long sequences from overlapping ESTs which are of sufficiently high quality to be able to unambiguously identify tags to individual members of multigene families, even when the sequences are highly conserved. A total of 106 genes encoding 50 different cytoplasmic ribosomal protein types have been identified, most proteins being encoded by at least two and up to four genes. Coding sequences of members of individual gene families are almost always very highly conserved and derived amino acid sequences are almost, if not completely, identical in the vast majority of cases. Sequence divergence is observed in untranslated regions which allows the definition of gene-specific probes. The method can be used to construct high-quality tags to any protein. 相似文献
997.
Brown RP 《Genetica》1997,101(1):67-74
Heterogeneous phenotypic correlations may be suggestive of underlying changes in genetic covariance among life-history, morphology,
and behavioural traits, and their detection is therefore relevant to many biological studies. Two new statistical tests are
proposed and their performances compared with existing methods. Of all tests considered, the existing approximate test of
homogeneity of product-moment correlations provides the greatest power to detect heterogeneous correlations, when based on
Hotelling's z*-transformation. The use of this transformation and test is recommended under conditions of bivariate normality.
A new distribution-free randomisation test of homogeneity of Spearman's rank correlations is described and recommended for
use when the bivariate samples are taken from populations with non-normal or unknown distributions. An alternative randomisation
test of homogeneity of product-moment correlations is shown to be a useful compromise between the approximate tests and the
randomisation tests on Spearman's rank correlations: it is not as sensitive to departures from normality as the approximate
tests, but has greater power than the rank correlation test. An example is provided that shows how choice of test will have
a considerable influence on the conclusions of a particular study.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
998.
999.
Dictyostelium RasG Is Required for Normal Motility and Cytokinesis, But Not Growth 总被引:1,自引:0,他引:1 下载免费PDF全文
Richard I. Tuxworth Janet L. Cheetham Laura M. Machesky George B. Spiegelmann Gerald Weeks Robert H. Insall 《The Journal of cell biology》1997,138(3):605-614
RasG is the most abundant Ras protein in growing Dictyostelium cells and the closest relative of mammalian Ras proteins. We have generated null mutants in which expression of RasG is completely abolished. Unexpectedly, RasG− cells are able to grow at nearly wild-type rates. However, they exhibit defective cell movement and a wide range of defects in the control of the actin cytoskeleton, including a loss of cell polarity, absence of normal lamellipodia, formation of unusual small, punctate polymerized actin structures, and a large number of abnormally long filopodia. Despite their lack of polarity and abnormal cytoskeleton, mutant cells perform normal chemotaxis. However, rasG− cells are unable to perform normal cytokinesis, becoming multinucleate when grown in suspension culture. Taken together, these data suggest a principal role for RasG in coordination of cell movement and control of the cytoskeleton. 相似文献
1000.
Genetic Analysis of Myoblast Fusion: blown fuse Is Required for Progression Beyond the Prefusion Complex 总被引:11,自引:1,他引:10 下载免费PDF全文
Stephen K. Doberstein Richard D. Fetter Anand Y. Mehta Corey S. Goodman 《The Journal of cell biology》1997,136(6):1249-1261
The events of myoblast fusion in Drosophila are dissected here by combining genetic analysis with light and electron microscopy. We describe a new and essential intermediate step in the process, the formation of a prefusion complex consisting of “paired vesicles.” These pairs of vesicles from different cells align with each other across apposed plasma membranes. This prefusion complex resolves into dense membrane plaques between apposed cells; these cells then establish cytoplasmic continuity by fusion of small areas of plasma membrane followed by vesiculation of apposed membranes. Different steps in this process are specifically blocked by mutations in four genes required for myoblast fusion. One of these genes, blown fuse, encodes a novel cytoplasmic protein expressed in unfused myoblasts that is essential for progression beyond the prefusion complex stage. 相似文献