全文获取类型
收费全文 | 1771篇 |
免费 | 101篇 |
国内免费 | 1篇 |
专业分类
1873篇 |
出版年
2022年 | 22篇 |
2021年 | 24篇 |
2020年 | 21篇 |
2019年 | 24篇 |
2018年 | 32篇 |
2017年 | 39篇 |
2016年 | 34篇 |
2015年 | 56篇 |
2014年 | 94篇 |
2013年 | 116篇 |
2012年 | 130篇 |
2011年 | 111篇 |
2010年 | 61篇 |
2009年 | 70篇 |
2008年 | 104篇 |
2007年 | 86篇 |
2006年 | 88篇 |
2005年 | 78篇 |
2004年 | 79篇 |
2003年 | 88篇 |
2002年 | 79篇 |
2001年 | 46篇 |
2000年 | 38篇 |
1999年 | 25篇 |
1998年 | 21篇 |
1997年 | 23篇 |
1996年 | 17篇 |
1995年 | 8篇 |
1994年 | 10篇 |
1993年 | 11篇 |
1992年 | 23篇 |
1991年 | 20篇 |
1990年 | 32篇 |
1989年 | 31篇 |
1988年 | 11篇 |
1987年 | 21篇 |
1986年 | 9篇 |
1985年 | 9篇 |
1984年 | 12篇 |
1983年 | 6篇 |
1982年 | 13篇 |
1981年 | 7篇 |
1980年 | 3篇 |
1979年 | 4篇 |
1978年 | 5篇 |
1975年 | 6篇 |
1971年 | 4篇 |
1968年 | 4篇 |
1967年 | 3篇 |
1959年 | 3篇 |
排序方式: 共有1873条查询结果,搜索用时 15 毫秒
31.
K Sato M Ishiai K Toda S Furukoshi A Osakabe H Tachiwana Y Takizawa W Kagawa H Kitao N Dohmae C Obuse H Kimura M Takata H Kurumizaka 《The EMBO journal》2012,31(17):3524-3536
Fanconi anaemia (FA) is a rare hereditary disorder characterized by genomic instability and cancer susceptibility. A key FA protein, FANCD2, is targeted to chromatin with its partner, FANCI, and plays a critical role in DNA crosslink repair. However, the molecular function of chromatin-bound FANCD2-FANCI is still poorly understood. In the present study, we found that FANCD2 possesses nucleosome-assembly activity in vitro. The mobility of histone H3 was reduced in FANCD2-knockdown cells following treatment with an interstrand DNA crosslinker, mitomycin C. Furthermore, cells harbouring FANCD2 mutations that were defective in nucleosome assembly displayed impaired survival upon cisplatin treatment. Although FANCI by itself lacked nucleosome-assembly activity, it significantly stimulated FANCD2-mediated nucleosome assembly. These observations suggest that FANCD2-FANCI may regulate chromatin dynamics during DNA repair. 相似文献
32.
Motoi Kanagawa Zhongpeng Lu Chiyomi Ito Chie Matsuda Katsuya Miyake Tatsushi Toda 《PloS one》2014,9(9)
Defects in dystroglycan glycosylation are associated with a group of muscular dystrophies, termed dystroglycanopathies, that include Fukuyama congenital muscular dystrophy (FCMD). It is widely believed that abnormal glycosylation of dystroglycan leads to disease-causing membrane fragility. We previously generated knock-in mice carrying a founder retrotransposal insertion in fukutin, the gene responsible for FCMD, but these mice did not develop muscular dystrophy, which hindered exploring therapeutic strategies. We hypothesized that dysferlin functions may contribute to muscle cell viability in the knock-in mice; however, pathological interactions between glycosylation abnormalities and dysferlin defects remain unexplored. To investigate contributions of dysferlin deficiency to the pathology of dystroglycanopathy, we have crossed dysferlin-deficient dysferlin
sjl/sjl mice to the fukutin-knock-in fukutin
Hp/− and Large-deficient Large
myd/myd mice, which are phenotypically distinct models of dystroglycanopathy. The fukutin
Hp/− mice do not show a dystrophic phenotype; however, (dysferlin
sjl/sjl: fukutin
Hp/−) mice showed a deteriorated phenotype compared with (dysferlin
sjl/sjl: fukutin
Hp/+) mice. These data indicate that the absence of functional dysferlin in the asymptomatic fukutin
Hp/− mice triggers disease manifestation and aggravates the dystrophic phenotype. A series of pathological analyses using double mutant mice for Large and dysferlin indicate that the protective effects of dysferlin appear diminished when the dystrophic pathology is severe and also may depend on the amount of dysferlin proteins. Together, our results show that dysferlin exerts protective effects on the fukutin
Hp/− FCMD mouse model, and the (dysferlin
sjl/sjl: fukutin
Hp/−) mice will be useful as a novel model for a recently proposed antisense oligonucleotide therapy for FCMD. 相似文献
33.
Covalent modifier NEDD8 is essential for SCF ubiquitin-ligase in fission yeast 总被引:6,自引:0,他引:6 下载免费PDF全文
Osaka F Saeki M Katayama S Aida N Toh-E A Kominami K Toda T Suzuki T Chiba T Tanaka K Kato S 《The EMBO journal》2000,19(13):3475-3484
A ubiquitin-like modifier, NEDD8, is covalently attached to cullin-family proteins, but its physiological role is poorly understood. Here we report that the NEDD8-modifying pathway is essential for cell viability and function of Pcu1 (cullin-1 orthologue) in fission yeast. Pcu1 assembled on SCF ubiquitin-ligase was completely modified by NEDD8. Pcu1(K713R) defective for NEDD8 conjugation lost the ability to complement lethality due to pcu1 deletion. Forced expression of Pcu1(K713R) or depletion of NEDD8 in cells resulted in impaired cell proliferation and marked stabilization of the cyclin-dependent kinase inhibitor Rum1, which is a substrate of the SCF complex. Based on these findings, we propose that covalent modification of cullin-1 by the NEDD8 system plays an essential role in the function of SCF in fission yeast. 相似文献
34.
35.
Functional Dissection and Hierarchy of Tubulin-folding Cofactor Homologues in Fission Yeast 总被引:2,自引:1,他引:2 下载免费PDF全文
Pippa A. Radcliffe Dai Hirata Leah Vardy Takashi Toda 《Molecular biology of the cell》1999,10(9):2987-3001
We describe the isolation of fission yeast homologues of tubulin-folding cofactors B (Alp11) and E (Alp21), which are essential for cell viability and the maintenance of microtubules. Alp11B contains the glycine-rich motif (the CLIP-170 domain) involved in microtubular functions, whereas, unlike mammalian cofactor E, Alp21E does not. Both mammalian and yeast cofactor E, however, do contain leucine-rich repeats. Immunoprecipitation analysis shows that Alp11B interacts with both α-tubulin and Alp21E, but not with the cofactor D homologue Alp1, whereas Alp21E also interacts with Alp1D. The cellular amount of α-tubulin is decreased in both alp1 and alp11 mutants. Overproduction of Alp11B results in cell lethality and the disappearance of microtubules, which is rescued by co-overproduction of α-tubulin. Both full-length Alp11B and the C-terminal third containing the CLIP-170 domain localize in the cytoplasm, and this domain is required for efficient binding to α-tubulin. Deletion of alp11 is suppressed by multicopy plasmids containing either alp21+ or alp1+, whereas alp21 deletion is rescued by overexpression of alp1+ but not alp11+. Finally, the alp1 mutant is not complemented by either alp11+ or alp21+. The results suggest that cofactors operate in a linear pathway (Alp11B-Alp21E-Alp1D), each with distinct roles. 相似文献
36.
37.
Mitsuru Kuwamura Yosuke Ando Akiko Takada Toshiko Kanehara Jyoji Yamate Takao Kotani Shigehito Takeshita Miyuki Kanbori Kazuhiro Kitada Tadao Serikawa 《Experimental Animals》2004,53(1):21-26
In this paper, we executed genome mapping and comparative mapping analyses for cvd and hob, autosomal recessive mutations with cerebellar vermis defect and cerebellar dysplasia in the rat. For the linkage analysis, we produced three sets of backcross progeny, (ACI x CVD)F(1) and (F344 x CVD)F(1) females crossed to a cvd homozygous male rat, and (HOB x WKY)F(1) males crossed to hob homozygous female rats. Analysis of the segregation patterns of simple sequence length polymorphism (SSLP) markers scanning the whole rat genome allowed the mapping of these autosomal recessive mutations to rat Chromosome (Chr) 2. The most likely gene order is D2Mgh12 - D2Rat86 - D2Mit15 - D2Rat185 - cvd - D2Rat66 - D2Mgh13, and D2Mit18 - Fga -D2Mit14 - D2Rat16 - hob - D2Mgh13. Crossing test between a proven cvd heterozygous and a hob heterozygous rats demonstrated their allelism. Furthermore, comparative mapping indicated the cvd locus corresponds to mouse chromosome 3 and a strong candidate gene Unc5h3, a causative gene for the rostral cerebellar malformation mouse, was implicated. 相似文献
38.
Cloning and sequence analysis of a cDNA encoding the Rieske iron-sulfur protein of rat mitochondrial cytochrome bc1 complex 总被引:2,自引:0,他引:2
M Nishikimi Y Hosokawa H Toda H Suzuki T Ozawa 《Biochemical and biophysical research communications》1989,159(1):19-25
We have isolated a cDNA clone for the Rieske iron-sulfur protein of rat cytochrome bc1 complex, by screening a rat liver cDNA expression library using antiserum directed against the corresponding protein of bovine. The amino acid sequence deduced from the nucleotide sequence of the cDNA indicated that the mature polypeptide of the rat protein consists of 196 amino acid residues with a molecular weight of 21,465, and that it is formed as a precursor with an amino-terminal extension. Northern blot analysis indicated that rat liver possibly contains different sizes of mRNAs for the Rieske iron-sulfur protein, and Southern blot analysis demonstrated that rats and mice possess a single gene for this protein. 相似文献
39.
Coomassie Brilliant Blue (CBB) protein stains are inexpensive but detect proteins at only at microgram levels. Because of acetic acid and methanol, they cause skin irritation and reduce work motivation by malodor. Recent mass spectrometric (MS) analyses demonstrated that nanogram-sensitive colloidal CBB staining resulted in in vitro methylations of proteins. We propose a rapid, inexpensive, sensitive, odorless, less harsh, and in vitro methylation-free CBB stain. CGP uses three components: citric acid, CBB G-250, and polyvinylpyrrolidone. CGP detects proteins at 12 ng within 45 min, and because it is nonalcohol, in principle in vitro methylation would be eliminated. Indeed, MS analysis of CGP-stained bands confirmed a lack of methylation. 相似文献
40.