全文获取类型
收费全文 | 2846篇 |
免费 | 197篇 |
国内免费 | 1篇 |
专业分类
3044篇 |
出版年
2023年 | 5篇 |
2022年 | 20篇 |
2021年 | 43篇 |
2020年 | 18篇 |
2019年 | 19篇 |
2018年 | 37篇 |
2017年 | 37篇 |
2016年 | 60篇 |
2015年 | 83篇 |
2014年 | 98篇 |
2013年 | 159篇 |
2012年 | 199篇 |
2011年 | 194篇 |
2010年 | 119篇 |
2009年 | 119篇 |
2008年 | 163篇 |
2007年 | 172篇 |
2006年 | 176篇 |
2005年 | 158篇 |
2004年 | 160篇 |
2003年 | 136篇 |
2002年 | 138篇 |
2001年 | 47篇 |
2000年 | 45篇 |
1999年 | 59篇 |
1998年 | 28篇 |
1997年 | 40篇 |
1996年 | 36篇 |
1995年 | 27篇 |
1994年 | 30篇 |
1993年 | 22篇 |
1992年 | 40篇 |
1991年 | 32篇 |
1990年 | 42篇 |
1989年 | 29篇 |
1988年 | 32篇 |
1987年 | 25篇 |
1986年 | 29篇 |
1985年 | 23篇 |
1984年 | 19篇 |
1983年 | 17篇 |
1982年 | 23篇 |
1981年 | 14篇 |
1980年 | 12篇 |
1979年 | 13篇 |
1978年 | 8篇 |
1977年 | 7篇 |
1975年 | 6篇 |
1973年 | 6篇 |
1968年 | 4篇 |
排序方式: 共有3044条查询结果,搜索用时 0 毫秒
51.
Watanabe M Yuzawa H Handa N Kobayashi I 《Applied and environmental microbiology》2006,72(8):5367-5375
Genome sequence comparisons among multiple species of Pyrococcus, a hyperthermophilic archaeon, revealed a linkage between a putative restriction-modification gene complex and several large genome polymorphisms/rearrangements. From a region apparently inserted into the Pyrococcus abyssi genome, a hyperthermoresistant restriction enzyme [PabI; 5'-(GTA/C)] with a novel structure was discovered. In the present work, the neighboring methyltransferase homologue, M.PabI, was characterized. Its N-terminal half showed high similarities to the M subunit of type I systems and a modification enzyme of an atypical type II system, M.AhdI, while its C-terminal half showed high similarity to the S subunit of type I systems. M.PabI expressed within Escherichia coli protected PabI sites from RsaI, a PabI isoschizomer. M.PabI, purified following overexpression, was shown to generate 5'-GTm6AC, which provides protection against PabI digestion. M.PabI was found to be highly thermophilic; it showed methylation at 95 degrees C and retained at least half the activity after 9 min at 95 degrees C. This hyperthermophilicity allowed us to obtain activation energy and other thermodynamic parameters for the first time for any DNA methyltransferases. We also determined the kinetic parameters of kcat, Km, DNA, and Km, AdoMet. The activity of M.PabI was optimal at a slightly acidic pH and at an NaCl concentration of 200 to 500 mM and was inhibited by Zn2+ but not by Mg2+, Ca2+, or Mn2+. These and previous results suggest that this unique methyltransferase and PabI constitute a type II restriction-modification gene complex that inserted into the P. abyssi genome relatively recently. As the most thermophilic of all the characterized DNA methyltransferases, M.PabI may help in the analysis of DNA methylation and its application to DNA engineering. 相似文献
52.
Futaki S Niwa M Nakase I Tadokoro A Zhang Y Nagaoka M Wakako N Sugiura Y 《Bioconjugate chemistry》2004,15(3):475-481
Arginine-rich peptide-mediated protein delivery into living cells is a novel technology for controlling cell functions with therapeutic potential. In this report, a novel approach for the intracellular delivery of histidine-tagged proteins was introduced where a Ni(II) chelate of octaarginine peptide bearing nitrilotriacetic acid [R8-NTA-Ni(II)] was used as a membrane-permeable carrier molecule. Significant internalization of histidine-tagged enhanced green fluorescent protein (EGFP) into HeLa cells was observed by confocal microscopic observation in the presence of R8-NTA-Ni(II). Nuclear condensation characteristic in apoptotic cell death was also induced in the cells treated with a histidine-tagged apoptosis-inducing peptide [pro-apoptotic domain peptide (PAD)], indicating that the cargo molecules really went through the membrane to reach the cytosol. The apoptosis-inducing activity of the peptide thus delivered was compared with that of the PAD peptide covalently connected with the octaarginine peptide. 相似文献
53.
Daichi Iwasaki Kayoko Hayashihara Hiroki Shima Mika Higashide Masahiro Terasawa Susan M. Gasser Miki Shinohara 《PLoS genetics》2016,12(3)
Because DNA double-strand breaks (DSBs) are one of the most cytotoxic DNA lesions and often cause genomic instability, precise repair of DSBs is vital for the maintenance of genomic stability. Xrs2/Nbs1 is a multi-functional regulatory subunit of the Mre11-Rad50-Xrs2/Nbs1 (MRX/N) complex, and its function is critical for the primary step of DSB repair, whether by homologous recombination (HR) or non-homologous end joining. In human NBS1, mutations result truncation of the N-terminus region, which contains a forkhead-associated (FHA) domain, cause Nijmegen breakage syndrome. Here we show that the Xrs2 FHA domain of budding yeast is required both to suppress the imprecise repair of DSBs and to promote the robust activation of Tel1 in the DNA damage response pathway. The role of the Xrs2 FHA domain in Tel1 activation was independent of the Tel1-binding activity of the Xrs2 C terminus, which mediates Tel1 recruitment to DSB ends. Both the Xrs2 FHA domain and Tel1 were required for the timely removal of the Ku complex from DSB ends, which correlates with a reduced frequency of imprecise end-joining. Thus, the Xrs2 FHA domain and Tel1 kinase work in a coordinated manner to maintain DSB repair fidelity. 相似文献
54.
55.
Chernyavsky AI Arredondo J Kitajima Y Sato-Nagai M Grando SA 《The Journal of biological chemistry》2007,282(18):13804-13812
Although it is accepted that pemphigus antibody binding to keratinocytes (KCs) evokes an array of intracellular biochemical events resulting in cell detachment and death, the triggering events remain obscure. It has been postulated that the binding of pemphigus vulgaris IgG (PVIgG) to KCs induces "desmosomal" signaling. Because in contrast to integrins and classical cadherins, desmoglein (Dsg) molecules are not known to elicit intracellular signaling, and because PV patients also produce non-Dsg autoantibodies, we investigated the roles of both Dsg and non-desmoglein PV antigens. The time course studies of KCs treated with PVIgG demonstrated that the activity of Src peaked at 30 min, EGF receptor kinase (EGFRK) at 60 min, and p38 MAPK at 240 min. The Src inhibitor PP2 decreased EGFRK and p38 activities by approximately 45 and 30%, respectively, indicating that in addition to Src, PVIgG evokes other triggering events. The shrinkage of KCs (cell volume reduction) became significant at 120 min, keratin aggregation at 240 min, and an increase of TUNEL positivity at 360 min. Pretreatment of KCs with PP2 blocked PVIgG-dependent cell shrinkage and keratin aggregation by approximately 50% and TUNEL positivity by approximately 25%. The p38 MAPK inhibitor PD169316 inhibited these effects by approximately 15, 20, and 70%, respectively. Transfection of KCs with small interfering RNAs that silenced expression of Dsg1 and/or Dsg3 proteins, blocked approximately 50% of p38 MAPK activity but did not significantly alter the PVIgG-dependent rise in Src and EGFRK activities. These results indicate that activation of p38 MAPK is a late signaling step associated with collapse of the cytoskeleton and disassembly of desmosomes caused by upstream events involving Src and EGFRK. Therefore, the early acantholytic events are triggered by non-Dsg antibodies. 相似文献
56.
Miki Takahasi Kazuo Okanoya 《Ethology : formerly Zeitschrift fur Tierpsychologie》2010,116(5):396-405
Song diversity results from the interactions between natural selection, sexual selection, and individual learning. To understand song diversity, all three factors must be considered collectively, not separately. Bengalese Finches were domesticated about 250 yr ago. Their courtship songs have become different from their ancestor, the White‐rumped Munia. Bengalese Finches sing songs with complex note‐to‐note transition patterns and with acoustically diverse song notes while White‐rumped Munias sing songs with fixed note sequence and mostly broad band song notes. Bengalese Finches were selected for domestication based on their good parenting ability, not their songs, but this artificial selection has nonetheless affected their songs. To test whether divergence occurred not only in the song phenotypes but also in the genetic basis for predisposition of strain specific song learning, we conducted a cross‐fostering experiment between Bengalese Finches and White‐rumped Munias. In both strains, song learning was affected by rearing condition: the acoustical feature and transition patterns followed those of the foster fathers. However, the accuracy of song learning differed between the wild and the domesticated strains: sharing of song note between sons and tutors in Finches was not very accurate regardless of the tutor, while Munias were highly accurate in copying Munia songs but often omitted song elements from Finch fathers. These results suggest that White‐rumped Munias are strongly constrained to learn their own strain’s song, and that this constraint was relaxed in the Bengalese Finch by domestication. 相似文献
57.
Kensuke Kobayashi Tomohiro Tsujita Hirokatsu Ito Satoshi Ozaki Takeshi Tani Yasuyuki Ishii Shoki Okuda Kiyoshi Tadano Takahiro Fukuroda Hisashi Ohta Osamu Okamoto 《Bioorganic & medicinal chemistry letters》2009,19(16):4729-4732
Structure–activity relationship studies directed toward improving the metabolic stability of compound 1 resulted in the identification of 3-[5-(3,5-difluorophenyl)-3-({[(1S,3R)-3-fluorocyclopentyl]amino}methyl)-4-methyl-1H-pyrazol-1-yl]propanenitrile 39 (MK-1925) as a selective, orally available and brain-penetrable opioid receptor-like 1 (ORL1) antagonist. The compound also showed in vivo efficacy after oral dosing. Therefore, compound 39 was selected to undergo further studies as a clinical candidate. 相似文献
58.
Takashi Hatakeyama Miki Okada Seiko Shimamoto Yasuo Kubota Ryoji Kobayashi 《European journal of biochemistry》2004,271(18):3765-3775
In this report, we have focused our attention on identifying intracellular mammalian proteins that bind S100A12 in a Ca2+-dependent manner. Using S100A12 affinity chromatography, we have identified cytosolic NADP+-dependent isocitrate dehydrogenase (IDH), fructose-1,6-bisphosphate aldolase A (aldolase), glyceraldehyde-3-phosphate dehydrogenese (GAPDH), annexin V, S100A9, and S100A12 itself as S100A12-binding proteins. Immunoprecipitation experiments indicated the formation of stable complexes between S100A12 and IDH, aldolase, GAPDH, annexin V and S100A9 in vivo. Surface plasmon resonance analysis showed that the binding to S100A12, of S100A12, S100A9 and annexin V, was strictly Ca2+-dependent, whereas that of GAPDH and IDH was only weakly Ca2+-dependent. To localize the site of S100A12 interaction, we examined the binding of a series of C-terminal truncation mutants to the S100A12-immobilized sensor chip. The results indicated that the S100A12-binding site on S100A12 itself is located at the C-terminus (residues 87-92). However, cross-linking experiments with the truncation mutants indicated that residues 87-92 were not essential for S100A12 dimerization. Thus, the interaction between S100A12 and S100A9 or immobilized S100A12 should not be viewed as a typical S100 homo- or heterodimerization model. Ca2+-dependent affinity chromatography revealed that C-terminal residues 75-92 are not necessary for the interaction of S100A12 with IDH, aldolase, GAPDH and annexin V. To analyze the functional properties of S100A12, we studied its action in protein folding reactions in vitro. The thermal aggregation of IDH or GAPDH was facilitated by S100A12 in the absence of Ca2+, whereas in the presence of Ca2+ the protein suppressed the aggregation of aldolase to less than 50%. These results suggest that S100A12 may have a chaperone/antichaperone-like function which is Ca2+-dependent. 相似文献
59.
60.