全文获取类型
收费全文 | 5183篇 |
免费 | 534篇 |
出版年
2023年 | 19篇 |
2022年 | 17篇 |
2021年 | 88篇 |
2020年 | 61篇 |
2019年 | 63篇 |
2018年 | 84篇 |
2017年 | 88篇 |
2016年 | 127篇 |
2015年 | 227篇 |
2014年 | 278篇 |
2013年 | 347篇 |
2012年 | 439篇 |
2011年 | 434篇 |
2010年 | 305篇 |
2009年 | 269篇 |
2008年 | 366篇 |
2007年 | 367篇 |
2006年 | 340篇 |
2005年 | 316篇 |
2004年 | 296篇 |
2003年 | 279篇 |
2002年 | 251篇 |
2001年 | 44篇 |
2000年 | 40篇 |
1999年 | 64篇 |
1998年 | 83篇 |
1997年 | 37篇 |
1996年 | 55篇 |
1995年 | 48篇 |
1994年 | 51篇 |
1993年 | 37篇 |
1992年 | 23篇 |
1991年 | 16篇 |
1990年 | 17篇 |
1989年 | 21篇 |
1988年 | 12篇 |
1987年 | 13篇 |
1986年 | 8篇 |
1985年 | 8篇 |
1984年 | 7篇 |
1983年 | 9篇 |
1982年 | 7篇 |
1981年 | 9篇 |
1980年 | 4篇 |
1979年 | 6篇 |
1978年 | 6篇 |
1977年 | 13篇 |
1976年 | 8篇 |
1974年 | 3篇 |
1972年 | 2篇 |
排序方式: 共有5717条查询结果,搜索用时 15 毫秒
21.
Alexander Schouten Jan Roosien Fred A. van Engelen G. A. M. de Jong A. W. M. Borst-Vrenssen Jacoline F. Zilverentant Dirk Bosch Willem J. Stiekema Fred J. Gommers Arjen Schots Jaap Bakker 《Plant molecular biology》1996,30(4):781-793
The effects of subcellular localization on single-chain antibody (scFv) expression levels in transgenic tobacco was evaluated using an scFv construct of a model antibody possessing different targeting signals. For translocation into the secretory pathway a secretory signal sequence preceded the scFv gene (scFv-S). For cytosolic expression the scFv antibody gene lacked such a signal sequence (scFv-C). Also, both constructs were provided with the endoplasmic reticulum (ER) retention signal KDEL (scFv-SK and scFv-CK, respectively). The expression of the different scFv constructs in transgenic tobacco plants was controlled by a CaMV 35S promoter with double enhancer. The scFv-S and scFv-SK antibody genes reached expression levels of 0.01% and 1% of the total soluble protein, respectively. Surprisingly, scFv-CK transformants showed considerable expression of up to 0.2% whereas scFv-C transformants did not show any accumulation of the scFv antibody. The differences in protein expression levels could not be explained by the steady-state levels of the mRNAs. Transient expression assays with leaf protoplasts confirmed these expression levels observed in transgenic plants, although the expression level of the scFv-S construct was higher. Furthermore, these assays showed that both the secretory signal and the ER retention signal were recognized in the plant cells. The scFv-CK protein was located intracellularly, presumably in the cytosol. The increase in scFv protein stability in the presence of the KDEL retention signal is discussed. 相似文献
22.
Hector M. Alvarez Frank Mayer Dirk Fabritius A. Steinbüchel 《Archives of microbiology》1996,165(6):377-386
An oleaginous hydrocarbon-degrading Rhodococcus opacus strain (PD630) was isolated from a soil sample. The cells were able to grow on a variety of substrates and to produce large
amounts of three different types of intracellular inclusions during growth on alkanes, phenylalkanes, or non-hydrocarbon substrates.
Electron microscopy revealed large numbers of electron-transparent inclusions with a sphere-like structure. In addition, electron-dense
inclusions representing polyphosphate and electron-transparent inclusions with an elongated disc-shaped morphology occurred
in small amounts. The electron-transparent inclusions of alkane- or gluconate-grown cells were composed of neutral lipids
(98%, w/w), phospholipids (1.2%, w/w), and protein (0.8%, w/w). The major component of the cellular inclusions was triacylglycerols;
minor amounts of diacylglycerols and probably also some free fatty acids were also present. Free fatty acids and/or fatty
acids in acylglycerols in cells of R. opacus amounted up to 76 or 87% of the cellular dry weight in gluconate- or olive-oil-grown cells, respectively. The fatty acid composition
of the inclusions depended on the substrate used for cultivation. In cells cultivated on n-alkanes, the composition of the
fatty acids was related to the substrate, and intermediates of the β-oxidation pathway, such as hexadecanoic or pentadecanoic
acid, were among the acylglycerols. Hexadecanoic acid was also the major fatty acid (up 36% of total fatty acids) occurring
in the lipid inclusions of gluconate-grown cells. This indicated that strain PD630 utilized β-oxidation and de novo fatty
acid biosynthesis for the synthesis of storage lipids. Inclusions isolated from phenyldecane-grown cells contained mainly
the non-modified substrate and phenylalkanoic acids derived from the hydrocarbon oxidation, such as phenyldecanoic acid, phenyloctanoic
acid, and phenylhexanoic acid, and approximately 5% (w/w) of diacylglycerols. The lipid inclusions seemed to have definite
structures, probably with membranes at their surfaces, which allow them to maintain their shape, and with some associated
proteins, probably involved in the inclusion formation.
Received: 22 December 1995 / Accepted: 12 March 1996 相似文献
23.
The nature of rhodopsin-triggered photocurrents in Chlamydomonas. II. Influence of monovalent ions. 总被引:1,自引:0,他引:1 下载免费PDF全文
Chlamydomonas exhibits a sequence of a photoreceptor current and two flagellar currents upon stimulation with bright green flashes. The currents are thought to be a prerequisite for the well-known photophobic responses. In the preceding paper, we analyzed the kinetics of these currents and their dependence on extracellular divalent ions. Here, we show that the photoreceptor current can be carried by monovalent ions (K+ > NH4+ > Na+), provided that the driving force is high enough. The small residual photoreceptor current observed in the absence of Ca2+ is able to evoke flagellar currents at low extracellular pH. This demonstrates that signal transduction from the rhodopsin to the flagella is not inevitably dependent on extracellular Ca2+. Double-flash experiments exclude a contribution of intra-rhodopsin charge movements to the photoreceptor current signal. Evidence will be provided for the existence of nonlocalized K+ outward currents, which counterbalance the localized Ca2+ influx and repolarize the cell after a light flash. A model is presented that explains the different pathways for direction changes and phobic responses. 相似文献
24.
Dirk Schories 《Aquatic Ecology》1995,29(3-4):341-347
For the last two decades dense mats of species of the filamentous green algaeEnteromorpha spp. have regulary occurred on tidal flats of Köningshafen Bay (island Sylt, North Sea, FRG). In calm areas overwintering of adult plants or plant fragments is a common process to guarantee the mass development during the next season. In contrast, the distribution ofEnteromorpha on exposed sandy tidal flats depends on recruitment by juvenile stages. In 1993Enteromorpha spore settlement was recorded regularly in the field. Polyethylene dishes were placed in the field and left for a period of seven days and lateron cultivated in the laboratory to checkEnteromorpha germling development. During summer 1993 — at a minimum distance of 200 m to the nearest adultEnteromorpha populations — a total of at least 82×106 spores m–2 settled. During winter the number of spores attached to the collecting dishes was close to zero and the adjacent sand flats were free of any visibleEnteromorpha plants. In further experiments it was shown that the development ofEnteromorpha juveniles in the next spring depended on the overwintering capacity of spores. More than 2×106 spores m–2 attached to large sand grains and other substrata (e.g. Hydrobia ulvae) survived the winter. In a laboratory experiment several species ofEnteromorpha were able to survive in total darkness for at least 10 months. 相似文献
25.
26.
27.
Insect resistance of transgenic plants that express modified Bacillus thuringiensis cryIA(b) and cryIC genes: a resistance management strategy 总被引:7,自引:0,他引:7
28.
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
29.
Lolium perenne growing with high root density on a fine nylon mesh (Kuchenbuch and Jungk, 1982) caused the development of element gradients in the rhizosphere below the mesh. Micro-liter soil solutions from 2-mg soil samples were sprayed onto Formvar-coated grids and analyzed by X-ray microanalysis in a transmission electron microscope. The results were comparable to those obtained by flame photometry and atomic absorption spectrometry (AAS) of conventional soil solutions from 1 g soil. X-ray microanalysis of micro-soil solutions allows the application of different extraction procedures to even small amounts of soil usually available from rhizosphere experiments. Information about soil buffering characteristics in the rhizosphere can thus be obtained. Aluminum accumulation in the rhizosphere of small segments of single Picea abies fine roots grown in undisturbed natural forest soil could be detected with this technique. 相似文献
30.
Three different stages of the avian schistosome Trichobilharziaocellata appeared to contain a novel N-acetylgalac-tosaminyltransferaseactivity. To investigate its function in the biosynthesis ofschistosome glycoconjugates, the enzyme was partially purifiedfrom cercariae, a free-living stage of the parasite, by affinitychromatography on UDP-Sepharose. Acceptor specificity studiesshowed that the enzyme catalyses the transfer of N-acetylgalactosamine(GalNAc) from UDP-GalNAc to oligosaccharides, glyco-peptidesand glycoproteins carrying a terminally 相似文献