全文获取类型
收费全文 | 43642篇 |
免费 | 3909篇 |
国内免费 | 33篇 |
专业分类
47584篇 |
出版年
2022年 | 315篇 |
2021年 | 604篇 |
2020年 | 397篇 |
2019年 | 492篇 |
2018年 | 645篇 |
2017年 | 562篇 |
2016年 | 1055篇 |
2015年 | 1767篇 |
2014年 | 1892篇 |
2013年 | 2471篇 |
2012年 | 3003篇 |
2011年 | 3090篇 |
2010年 | 1963篇 |
2009年 | 1669篇 |
2008年 | 2538篇 |
2007年 | 2524篇 |
2006年 | 2292篇 |
2005年 | 2356篇 |
2004年 | 2309篇 |
2003年 | 2203篇 |
2002年 | 2157篇 |
2001年 | 522篇 |
2000年 | 426篇 |
1999年 | 533篇 |
1998年 | 615篇 |
1997年 | 467篇 |
1996年 | 416篇 |
1995年 | 399篇 |
1994年 | 359篇 |
1993年 | 353篇 |
1992年 | 397篇 |
1991年 | 336篇 |
1990年 | 272篇 |
1989年 | 281篇 |
1988年 | 283篇 |
1987年 | 256篇 |
1986年 | 239篇 |
1985年 | 310篇 |
1984年 | 326篇 |
1983年 | 262篇 |
1982年 | 354篇 |
1981年 | 278篇 |
1980年 | 259篇 |
1979年 | 194篇 |
1978年 | 223篇 |
1977年 | 213篇 |
1976年 | 181篇 |
1975年 | 172篇 |
1974年 | 196篇 |
1973年 | 200篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
241.
Structural and enzymatic studies of the T4 DNA replication system. I. Physical characterization of the polymerase accessory protein complex 总被引:10,自引:0,他引:10
In this study, we have investigated the structural and physical properties of the bacteriophage T4 DNA polymerase accessory proteins. We find that T4 gene 44 and 62 proteins associate to form a tight, highly homogeneous complex, containing four gene 44 protein subunits and one gene 62 protein subunit. The molecular mass of the complex is 163,700 daltons. Sedimentation results suggest that the complex is quite asymmetric, with a prolate ellipsoid axial ratio of about 5:1. This protein complex is known to carry a DNA-dependent ATPase activity; we show by photoaffinity labeling that the ATP-binding sites reside in the gene 44 protein subunits of the complex. Equilibrium sedimentation and chemical cross-linking studies indicate that the T4 gene 45 protein self-associates to form a trimer in solution. This trimer species also appears to be quite asymmetric, showing an axial ratio for a prolate ellipsoid of about 6:1, assuming normal hydration. 相似文献
242.
243.
Wayne W. Grody Deborah Klein Amy E. Dodson Rita M. Kern Paul B. Wissmann Barbara K. Goodman Patrick Bassand Bert Marescau Soo-Sang Kang James V. Leonard Stephen D. Cederbaum 《American journal of human genetics》1992,50(6):1281-1290
We have explored the molecular pathology in 28 individuals homozygous or heterozygous for liver arginase deficiency (hyperargininemia) by a combination of Southern analysis, western blotting, DNA sequencing, and PCR. This cohort represents the majority of arginase-deficient individuals worldwide. Only 2 of 15 homozygous patients on whom red blood cells were available had antigenically cross-reacting material as ascertained by western blot analysis using anti-liver arginase antibody. Southern blots of patient genomic DNAs, cut with a variety of restriction enzymes and probed with a near-full-length (1,450-bp) human liver arginase cDNA clone, detected no gross gene deletions. Loss of a TaqI cleavage site was identified in three individuals: in a homozygous state in a Saudi Arabian patient at one site, at a different site in homozygosity in a German patient, and in heterozygosity in a patient from Australia. The changes in the latter two were localized to exon 8, through amplification of this region by PCR and electrophoretic analysis of the amplified fragment after treatment with TaqI; the precise base changes (Arg291X and Thr290Ser) were confirmed by sequencing. It is interesting that the latter nucleotide variant (Thr290Ser) was found to lie adjacent to the TaqI site rather than within it, though whether such a conservative amino acid substitution represents a true pathologic mutation remains to be determined. We conclude that arginase deficiency, though rare, is a heterogeneous disorder at the genotypic level, generally encompassing a variety of point mutations rather than substantial structural gene deletions. 相似文献
244.
A stable xylan suspension was prepared and characterized. Hydrolysis of the particles converts them into soluble fragments, thereby lowering the turbidity of the suspension. The small volume of the assay mixture, the short incubation time required, and the simplicity of the procedure permit the rapid analysis of many samples. Furthermore, the procedure can be used to assay xylanase activities in the presence of other reducing materials and is also useful for monitoring low-level xylanase activities. 相似文献
245.
B4-2-1 cells (Lec15 cells) are Chinese hamster ovary cells deficient in mannosylphosphoryldolichol synthase activity. They synthesize the truncated lipid intermediate Man5GlcNAc2-P-P-dolichol rather than the Glc3Man9GlcNAc2-P-P-dolichol synthesized by wild-type cells. In this report we present evidence that these cells did synthesize glucosylated Man5GlcNAc2-P-P-dolichol, but this species represented only a minor fraction of the labeled oligosaccharide-lipid. On the other hand, glucosylated oligosaccharides were a major species transferred to protein in these cells, showing that in vivo, glucosylated oligosaccharides are preferentially transferred to protein. The truncated oligosaccharides found in B4-2-1 cells were removed from the protein by N-glycanase treatment, since they were resistant to both endo-beta-N-acetylglucosaminidase H and F activity. B4-2-1 cells processed the glucosylated, truncated oligosaccharides transferred to G protein of vesicular stomatitis virus, leading to infectious virus. 相似文献
246.
Summary ABacillus sp. screened from termite infested soils produced significant amount of endoglucanase and xylanase enzymes when grown on a lignocellulosic substrate, rice husk. Biosynthesis of these enzymes was significantly enhanced by the addition of 0.2% cellobiose or glucose for endoglucanase and xylose for -xylanase activities. In the actual hydrolyses, glucose and cellobiose at low concentrations acted as activitors of endoglucanase activity whereas cellobiose and xylose acted as inhibitors of -xylanase activity. 相似文献
247.
Characterization of a pollen-specific gene family from Brassica napus which is activated during early microspore development 总被引:13,自引:0,他引:13
Diego Albani Laurian S. Robert Pauline A. Donaldson Illimar Altosaar Paul G. Arnison Steven F. Fabijanski 《Plant molecular biology》1990,15(4):605-622
In this paper we describe the isolation and characterization of a genomic clone (Bp4) from Brassica napus which contains three members of a pollen-specific multigene family. This family is composed of 10 to 15 closely related genes which are expressed in early stages of microspore development. The complete nucleotide sequence of the clone Bp4 and of three homologous cDNA clones is reported. One of the genes (Bp4B) contained in the genomic clone is believed to be non-functional because of sequence rearrangements in its 5 region and intron splicing sites. The remaining genes (Bp4A and Bp4C), as well as the cDNA clones, appear to code for small proteins of unique structure. Three different types of proteins can be predicted as a result of the deletion of carboxy or amino terminal portions of a conserved core protein. These proteins all share a common alternation of hydrophobic and hydrophilic domains. A fragment of the genomic clone containing the gene Bp4A, as well as the non-functional gene Bp4B, was introduced into tobacco plants via Agrobacterium-mediated transformation. The functional gene Bp4A is expressed in transgenic tobacco plants and shows spatial and temporal regulation consistent with the expression patterns seen in Brassica napus. 相似文献
248.
Paul G. Arnison Pauline Donaldson Anne Jackson Charmaine Semple Wilf Keller 《Plant Cell, Tissue and Organ Culture》1990,20(3):217-222
Anther culture medium was prepared with different types and concentrations of cytokinins to gain greater insight into the control of embryo formation during Brassica oleracea L. var. italica (broccoli) anther culture. The independent addition of the four cytokinins tested had widely divergent effects dependent upon cytokinin concentration and the genetic background of the test plants. All cytokinins were generally inhibitory at high concentrations, however, individual plants showed significant stimulation of embyro formation at typical physiological levels. The influence of cytokinins was highly cultivar-specific, some lines were stimulated, others inhibited and still other test lines were largely unaffected. Although the addition of cytokinins was needed for embryo formation for some plants, in no instance were cytokinins able to replace the inductive effect of high-temperature treatments. 相似文献
249.
Cora-Jean S. Edgell Jill E. Haizlip C. Robert Bagnell Joan P. Packenham Paul Harrison Barry Wilbourn Victoria J. Madden 《In vitro cellular & developmental biology. Plant》1990,26(12):1167-1172
Summary Weibel-Palade bodies are ultrastructurally defined organelles found only in vascular endothelial cells. Because endothelium
in corpo is very dispersed, isolation and further characterization of this organelle has been dependent on increasing the
number of cells in culture. However, primary isolates of endothelial cells have a limited replication potential and tend to
senesce in culture. In this report, EA.hy926, a continuously replicating cell line derived from human endothelium, is shown
to contain Weibel-Palade bodies. Electron micrographs demonstrate the ultrastructural characteristics of these tissue-specific
organelles and their cytoplasmic distribution in EA.hy926 cells. Von Willebrand factor, which has been shown to exist in Weibel
Palade bodies, is demonstrated by immunofluorescence in discrete rod-shaped organelles whose size, shape, and distribution
are consistent with that of Weibel-Palade bodies in primary endothelial cell cultures. Rapid release of von Willebrand factor
can be induced by calcium ionophore, and large multimeric forms of the protein are found in EA.hy926 cells. These two properties
are consistent with the function currently ascribed to Weibel Palade bodies: storage of multimerized von Willebrand factor.
Thus ultrastructural, immunologic, and functional data establish the existence of this as yet poorly understood tissue-specific
organelle in a continuous, vigorously replicating human cell line. 相似文献
250.
Cytochalasin B (CB) is known to interfere reversibly with the cytoplasmic contractile filamental network of mammalian cells. The role of the microfilament system in the mechanism of the reactive oxygen intermediates release of polymorphonuclear leukocytes (PMNL) was studied for different kinds of stimuli. PMNL from fresh human blood were treated with CB and stimulated by adherence on plastic surfaces, by opsonized zymosan, by phorbol myristate acetate and by N-formylmethionyl-phenylalaline. The production of reactive oxygen species were monitored by simultaneous detection of native, luminol-independent, luminescence (NL) and luminol-dependent luminescence (LDL) using a method of spectral discrimination. Different influences of CB on NL with respect to LDL as well stimuli-dependent influences of CB on the luminescence response of PMNL were observed. Especially phagocytosis-associated activation of PMNL was strongly inhibited by CB, whereas LDL was reduced to a much greater extent in comparison with NL. A firm involvement of the microfilament system is indicated, but it depends on the kind of stimulus engaged. 相似文献