全文获取类型
收费全文 | 11197篇 |
免费 | 1263篇 |
国内免费 | 1173篇 |
出版年
2024年 | 12篇 |
2023年 | 138篇 |
2022年 | 234篇 |
2021年 | 571篇 |
2020年 | 500篇 |
2019年 | 543篇 |
2018年 | 566篇 |
2017年 | 429篇 |
2016年 | 535篇 |
2015年 | 766篇 |
2014年 | 871篇 |
2013年 | 864篇 |
2012年 | 993篇 |
2011年 | 889篇 |
2010年 | 617篇 |
2009年 | 503篇 |
2008年 | 605篇 |
2007年 | 527篇 |
2006年 | 479篇 |
2005年 | 376篇 |
2004年 | 387篇 |
2003年 | 381篇 |
2002年 | 366篇 |
2001年 | 244篇 |
2000年 | 163篇 |
1999年 | 200篇 |
1998年 | 112篇 |
1997年 | 99篇 |
1996年 | 97篇 |
1995年 | 98篇 |
1994年 | 71篇 |
1993年 | 56篇 |
1992年 | 55篇 |
1991年 | 48篇 |
1990年 | 43篇 |
1989年 | 43篇 |
1988年 | 30篇 |
1987年 | 21篇 |
1986年 | 18篇 |
1985年 | 9篇 |
1984年 | 12篇 |
1983年 | 14篇 |
1982年 | 13篇 |
1981年 | 10篇 |
1980年 | 6篇 |
1979年 | 6篇 |
1977年 | 2篇 |
1975年 | 2篇 |
1974年 | 2篇 |
1972年 | 2篇 |
排序方式: 共有10000条查询结果,搜索用时 187 毫秒
991.
Candida tropicalis isolated from acclimated activated sludge was used in this study. Cell suspensions with 5 x 10(7) cells ml(-1) were irradiated by using a He-Ne laser. After mutagenesis, the irradiated cell suspension was diluted and plated on yeast extract-peptone-dextrose (YEPD) medium. Plates with approximately 20 individual colonies were selected, and all individual colonies were harvested for phenol biodegradation. The phenol biodegradation stabilities for 70 phenol biodegradation-positive mutants, mutant strains CTM 1 to 70, ranked according to their original phenol biodegradation potentials, were tested continuously during transfers. Finally, mutant strain CTM 2, which degraded 2,600 mg liter(-1) phenol within 70.5 h, was obtained on the basis of its capacity and hereditary stability for phenol biodegradation. The phenol hydroxylase gene sequences were cloned in wild and mutant strains. The results showed that four amino acids were mutated by irradiation with a laser. In order to compare the activity of phenol hydroxylase in wild and mutant strains, their genes were expressed in Escherichia coli BL21(DE3) and enzyme activities were spectrophotometrically determined. It was clear that the activity of phenol hydroxylase was promoted after irradiation with a He-Ne laser. In addition, the cell growth and intrinsic phenol biodegradation kinetics of mutant strain CTM 2 in batch cultures were also described by Haldane's kinetic equation with a wide range of initial phenol concentrations from 0 to 2,600 mg liter(-1). The specific growth and degradation rates further demonstrated that the CTM 2 mutant strain possessed a higher capacity to resist phenol toxicity than wild C. tropicalis did. 相似文献
992.
Fieseler L Hentschel U Grozdanov L Schirmer A Wen G Platzer M Hrvatin S Butzke D Zimmermann K Piel J 《Applied and environmental microbiology》2007,73(7):2144-2155
Numerous marine sponges harbor enormous amounts of as-yet-uncultivated bacteria in their tissues. There is increasing evidence that these symbionts play an important role in the synthesis of protective metabolites, many of which are of great pharmacological interest. In this study, genes for the biosynthesis of polyketides, one of the most important classes of bioactive natural products, were systematically investigated in 20 demosponge species from different oceans. Unexpectedly, the sponge metagenomes were dominated by a ubiquitously present, evolutionarily distinct, and highly sponge-specific group of polyketide synthases (PKSs). Open reading frames resembling animal fatty acid genes were found on three corresponding DNA regions isolated from the metagenomes of Theonella swinhoei and Aplysina aerophoba. Their architecture suggests that methyl-branched fatty acids are the metabolic product. According to a phylogenetic analysis of housekeeping genes, at least one of the PKSs belongs to a bacterium of the Deinococcus-Thermus phylum. The results provide new insights into the chemistry of sponge symbionts and allow inference of a detailed phylogeny of the diverse functional PKS types present in sponge metagenomes. Based on these qualitative and quantitative data, we propose a significantly simplified strategy for the targeted isolation of biomedically relevant PKS genes from complex sponge-symbiont associations. 相似文献
993.
Wen Liu Xuesen Chen Guanjun Liu Qing Liang Tianming He Jianrong Feng 《Plant Cell, Tissue and Organ Culture》2007,88(3):289-299
Embryo rescue technique was used successfully to produce interspecific hybrids by crossing peach (P. persica) as a female parent with apricot (P. armeniaca) and plum (P. salicica). In those crosses that had ‘Yuhualu’ or ‘Zhonghuashoutao’ as female parents, hybrid embryos aborted from the 7th or 8th
week after pollination mainly due to post-pollination incompatibility. An embryo rescue protocol was established to rescue
such embryos and recover hybrid plants. Modified half-strength MS medium containing 4 mg l−1 6-BA and 0.5 mg l−1 IBA produced up to 90% germination in the embryos. Modified MS medium with 1.0 mg l−1 6-BA and 1.0 mg l−1 IBA gave the highest bud induction and multiplication whereas modified MS medium containing 0.5 mg l−1 IAA and 0.2 mg l−1 NAA gave the best rooting percentage. All the hybrids obtained using this embryo rescue technique were verified using simple
sequence repeat (SSR) markers. A series of pollen treatments were carried out to partially overcome pre-pollination incompatibility,
and it was found accidentally that pollen treatment with electrostatic field not only improved pollen germination but also
increased the multiplication coefficient of embryo-induced shoots. 相似文献
994.
Two novel loci for pollen sterility in hybrids between the weedy strain Ludao and the Japonica variety Akihikari of rice (Oryza sativa L.) 总被引:4,自引:0,他引:4
Jing W Zhang W Jiang L Chen L Zhai H Wan J 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2007,114(5):915-925
Partial pollen sterility has been observed in hybrid progeny derived from a japonica cultivar, Akihikari and a weedy strain, Ludao, which naturally grows in Jiangsu province of east China. Cytological and histological
analyses revealed that pollen abortion occurred largely at the bicellular pollen stage, primarily due to the gradual disaggregation
of generative and vegetative cells. A genome-wide analysis was further carried out in a backcross population of Akihikari
//Ludao/Akihikari using a total of 118 simple sequence repeat (SSR) markers and an expressed sequence tag (EST) marker distributed
on the entire rice linkage map. Two loci controlling hybrid pollen sterility, designated as S33(t) and S34(t), were located on chromosomes 3 and 11, respectively. Both loci were putatively different from all the previously reported
gametophyte genes and hybrid pollen sterility loci. Interaction between the Ludao and Akihikari alleles at each of the two
loci resulted in reduction of fertility in the pollens carring the Ludao alleles. To map the precise location of the major
locus, S33(t), we selected 165 plants of the backcross population with pollen fertility higher than 80.0%, and assayed the recombinant
events surrounding the S33(t) locus using newly developed SSR markers. The S33(t) was delimited to an 86 kb region between SSR markers RM15621 and RM15627. Sequence analysis of this region indicated that
there were ten open reading frames. These results will be valuable for cloning this gene and marker-assisted transferring
of the corresponding neutral allele in rice breeding programs. Furthermore, the origin of the weedy strain Ludao is discussed. 相似文献
995.
This work was to characterize the generation of nitric oxide (NO) in Taxus yunnanensis cells induced by a fungal-derived cerebroside and the signal role of NO in the elicitation of plant defense responses and
taxol production. (2S,2′R,3R,3′E,4E,8E)-1-O-β-d-glucopyranosyl-2-N-(2′-hydroxy-3′-octadecenoyl)-3-hydroxy-9-methyl-4,8-sphingadienine at 10 μg/ml induced a rapid and dose-dependent NO production
in the Taxus cell culture, reaching a maximum within 5 h of the treatment. The NO donor sodium nitroprusside (SNP) potentiated cerebroside-induced
H2O2 production and cell death. Inhibition of nitric oxide synthase activity by phenylene-1,3-bis(ethane-2-isothiourea) dihydrobromide
or scavenging NO by 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide partially blocked the cerebroside-induced
H2O2 production and cell death. Moreover, NO enhanced cerebroside-induced activation of phenylalanine ammonium-lyase and accumulation
of taxol in cell cultures. These results are suggestive of a role for NO as a new signal component for activating the cerebroside-induced
defense responses and secondary metabolism activities of plant cells.
Taxol is a trademark of Bristol-Myers Squibb, Madison, NJ. 相似文献
996.
Jiang Q Huang J Li WQ Cavinato T Keller JR Durum SK 《Journal of immunology (Baltimore, Md. : 1950)》2007,178(1):228-234
Signals from the IL-7R are uniquely required for T cell development and maintenance, despite the resemblance of IL-7R to other cytokine receptors and the apparent sharing of common signaling pathways. This unique requirement could either reflect unique expression of IL-7R or IL-7, or it could indicate that the IL-7R delivers unique signals. To determine whether the IL-7R provided unique signals, we exchanged its intracellular domain with that of other cytokine receptors: IL-4R, IL-9R, and prolactin receptor (PRLR). Chimeric receptors were used to reconstitute development of IL-7R(-/-) hemopoietic progenitors by transducing the receptors in retroviral vectors. Whereas IL-7R(-/-) thymocytes are arrested at the double-negative stage, IL-4R, IL-9R, or PRLR all imparted some progression to the double-positive stage. IL-4R and PRLR gave only small numbers of thymocytes, whereas IL-9R gave robust alphabeta T cell development and reconstitution of peripheral CD4 and CD8 cells, indicating that it can duplicate many of the functions of IL-7R. However, IL-9R failed to reconstitute rearrangement of the TCRgamma locus or development of gammadelta T cells. Thus, the IL-7R signals required in the alphabeta T cell lineage (such as survival and proliferation) are not unique to this receptor, whereas rearrangement of the TCRgamma locus may require a signal that is not shared by other receptors. 相似文献
997.
Chen Y Pappu BP Zeng H Xue L Morris SW Lin X Wen R Wang D 《Journal of immunology (Baltimore, Md. : 1950)》2007,178(1):49-57
The adaptor protein B cell lymphoma 10 (Bcl10) plays an essential role in the functions of the AgRs in T and B cells. In this study, we report that Bcl10 also plays an important role in mast cells. Bcl10 is expressed in mast cells. Although Bcl10-deficient mast cells undergo normal development, we demonstrate that Bcl10 is essential for specific functions of FcepsilonR. Although Bcl10-deficient mast cells have normal de novo synthesis and release of the lipid mediator arachidonic acid, the mutant cells possess impaired FcepsilonR-mediated degranulation, indicated by decreased serotonin release, and impaired cytokine production, measured by release of IL-6. In addition, Bcl10-deficient mice display impaired IgE-mediated passive cutaneous anaphylaxis. Moreover, although Bcl10-deficient mast cells have normal FcepsilonR-mediated Ca(2+) flux, activation of PI3K, and activation of the three types of MAPKs (ERKs, JNK, and p38), the mutant cells have markedly diminished FcepsilonR-mediated activation of NF-kappaB and decreased activation of AP-1. Thus, Bcl10 is essential for FcepsilonR-induced activation of AP-1, NF-kappaB, degranulation, and cytokine production in mast cells. 相似文献
998.
Damaj BB Becerra CB Esber HJ Wen Y Maghazachi AA 《Journal of immunology (Baltimore, Md. : 1950)》2007,179(11):7907-7915
We describe here the protein expression of H4 histamine receptor in cells of the innate immune system, which include NK cells, monocytes, and dendritic cells (DCs). Anti-H4R specifically stained permeabilized NK cells, THP-1 clone 15 monocytes, and DCs. This binding was inhibited by incubating anti-H4R Ab with its corresponding peptide. Histamine induced NK cells, THP-1 clone 15 cells, and DCs chemotaxis with high affinity. The ED(50) chemotactic effect was 5 nM, 6.8 nM, and 2.7 nM for NK cells, THP-1 clone 15 cells, and DCs, respectively. Thioperamide, an H3R/H4R antagonist, inhibited histamine-induced chemotaxis in all these cells. However, histamine failed to induce the mobilization of [Ca(2+)](i) in NK cells and THP-1 clone 15 cells, but it induced calcium fluxes in DCs. Using a new method of detecting NK cell-mediated cytolysis, it was observed that NK cells efficiently lysed K562 target cells and that histamine did not affect this NK cell activity. In summary, this is the first demonstration of the protein expression of H4 receptor in NK cells. Also, the results of the chemotactic effects of histamine on NK cells and THP-1 cells are novel. These results may shed some light on the colocalization of cells of innate immune arm at sites of inflammation. They are also important for developing drugs that target H4R for the treatment of various disorders, such as autoimmune and immunodeficient diseases. 相似文献
999.
Pope WH Weigele PR Chang J Pedulla ML Ford ME Houtz JM Jiang W Chiu W Hatfull GF Hendrix RW King J 《Journal of molecular biology》2007,368(4):966-981
Marine Synechococcus spp and marine Prochlorococcus spp are numerically dominant photoautotrophs in the open oceans and contributors to the global carbon cycle. Syn5 is a short-tailed cyanophage isolated from the Sargasso Sea on Synechococcus strain WH8109. Syn5 has been grown in WH8109 to high titer in the laboratory and purified and concentrated retaining infectivity. Genome sequencing and annotation of Syn5 revealed that the linear genome is 46,214 bp with a 237 bp terminal direct repeat. Sixty-one open reading frames (ORFs) were identified. Based on genomic organization and sequence similarity to known protein sequences within GenBank, Syn5 shares features with T7-like phages. The presence of a putative integrase suggests access to a temperate life cycle. Assignment of 11 ORFs to structural proteins found within the phage virion was confirmed by mass-spectrometry and N-terminal sequencing. Eight of these identified structural proteins exhibited amino acid sequence similarity to enteric phage proteins. The remaining three virion proteins did not resemble any known phage sequences in GenBank as of August 2006. Cryo-electron micrographs of purified Syn5 virions revealed that the capsid has a single “horn”, a novel fibrous structure protruding from the opposing end of the capsid from the tail of the virion. The tail appendage displayed an apparent 3-fold rather than 6-fold symmetry. An 18 Å resolution icosahedral reconstruction of the capsid revealed a T = 7 lattice, but with an unusual pattern of surface knobs. This phage/host system should allow detailed investigation of the physiology and biochemistry of phage propagation in marine photosynthetic bacteria. 相似文献
1000.
Epstein-Barr virus BZLF1 gene, a switch from latency to lytic infection, is expressed as an immediate-early gene after primary infection of B lymphocytes 下载免费PDF全文
We demonstrate here that the Epstein-Barr virus (EBV) BZLF1 gene, a switch from latent infection to lytic infection, is expressed as early as 1.5 h after EBV infection in Burkitt's lymphoma-derived, EBV-negative Akata and Daudi cells and primary B lymphocytes. Since BZLF1 mRNA is expressed even when the cells are infected with EBV in the presence of anisomycin, an inhibitor of protein synthesis, its expression does not require prerequisite protein synthesis, indicating that BZLF1 is expressed as an immediate-early gene following primary EBV infection of B lymphocytes. 相似文献