全文获取类型
收费全文 | 1258篇 |
免费 | 80篇 |
国内免费 | 1篇 |
专业分类
1339篇 |
出版年
2022年 | 12篇 |
2021年 | 20篇 |
2020年 | 15篇 |
2019年 | 18篇 |
2018年 | 22篇 |
2017年 | 18篇 |
2016年 | 38篇 |
2015年 | 59篇 |
2014年 | 58篇 |
2013年 | 76篇 |
2012年 | 93篇 |
2011年 | 65篇 |
2010年 | 37篇 |
2009年 | 50篇 |
2008年 | 60篇 |
2007年 | 60篇 |
2006年 | 64篇 |
2005年 | 50篇 |
2004年 | 50篇 |
2003年 | 41篇 |
2002年 | 48篇 |
2001年 | 6篇 |
2000年 | 9篇 |
1999年 | 15篇 |
1998年 | 22篇 |
1997年 | 17篇 |
1996年 | 15篇 |
1995年 | 21篇 |
1994年 | 20篇 |
1993年 | 12篇 |
1992年 | 20篇 |
1991年 | 11篇 |
1990年 | 15篇 |
1989年 | 8篇 |
1988年 | 5篇 |
1984年 | 6篇 |
1983年 | 5篇 |
1982年 | 12篇 |
1981年 | 18篇 |
1980年 | 10篇 |
1979年 | 11篇 |
1978年 | 9篇 |
1977年 | 11篇 |
1976年 | 12篇 |
1975年 | 14篇 |
1974年 | 12篇 |
1973年 | 7篇 |
1971年 | 8篇 |
1970年 | 5篇 |
1967年 | 8篇 |
排序方式: 共有1339条查询结果,搜索用时 14 毫秒
31.
Kristel Eggermont Inge J. Goderis Willem F. Broekaert 《Plant Molecular Biology Reporter》1996,14(3):273-279
We describe here a reliable high-throughput method for extraction of RNA from fresh or frozen plant tissue that obviates laborious
and time-consuming homogenisation by mortar and pestle. The method is based on homogenisation by high-speed reciprocal shaking
in presence of a mixture of inexpensive abrasive materials; i.e., quartz sand and glass beads. After homogenisation, the method
follows a standard procedure for RNA extraction by phenol/LiCl. Yield and quality of RNA obtained by homogenisation with the
sand/glass bead mix are identical to those obtained by mortar and pestle. 相似文献
32.
Korzelius J The I Ruijtenberg S Portegijs V Xu H Horvitz HR van den Heuvel S 《Developmental biology》2011,(2):358-369
DNA replication and its connection to M phase restraint are studied extensively at the level of single cells but rarely in the context of a developing animal. C. elegans lin-6 mutants lack DNA synthesis in postembryonic somatic cell lineages, while entry into mitosis continues. These mutants grow slowly and either die during larval development or develop into sterile adults. We found that lin-6 corresponds to mcm-4 and encodes an evolutionarily conserved component of the MCM2-7 pre-RC and replicative helicase complex. The MCM-4 protein is expressed in all dividing cells during embryonic and postembryonic development and associates with chromatin in late anaphase. Induction of cell cycle entry and differentiation continues in developing mcm-4 larvae, even in cells that went through abortive division. In contrast to somatic cells in mcm-4 mutants, the gonad continues DNA replication and cell division until late larval development. Expression of MCM-4 in the epidermis (also known as hypodermis) is sufficient to rescue the growth retardation and lethality of mcm-4 mutants. While the somatic gonad and germline show substantial ability to cope with lack of zygotic mcm-4 function, mcm-4 is specifically required in the epidermis for growth and survival of the whole organism. Thus, C. elegans mcm-4 has conserved functions in DNA replication and replication checkpoint control but also shows unexpected tissue-specific requirements. 相似文献
33.
Dr. Inge Kayser-Wegmann 《Cell and tissue research》1976,171(4):513-521
Summary Black cuticles of larvae and pupae from various Lepidoptera were studied by light and electron microscopy. There are striking differences in the representation of black pigmentation, especially at the ultrastructural level. Two types may be described: 1. With the light microscope black melanin-like grana, electron-dense electron microscopically, are found in the distal parts of the exocuticle. This type is demonstrated in larvae of Celerio euphorbiae, Papilio machaon, and Phalera bucephala. 2. With the light microscope a dark homogeneous layer in the distal exocuticle can be recognized, however, electron microscopically no structures correlated with this dark pigment layer. This type of pigmentation was present in pupae of Pieris brassicae and Aglais urticae; in Pieris larvae the dark pigmented layer appeared to be limited to the epicuticle. In Celerio processes of the epidermal cells are involved in transporting precursors to the exocuticle. The conclusion was reached that black pigmentation in cuticles is based on different mechanisms as proposed by structural features. The two likely mechanisms are melanization and sclerotization.This work was supported by the Deutsche Forschungsgemeinschaft (SFB 87, project A1, granted to Prof. Bückmann) 相似文献
34.
Filamentous fungi are capable of secreting relatively large amounts of heterologous recombinant proteins. Recombinant human glycoproteins expressed in this system, however, carry only carbohydrates of the oligomannose type limiting their potential use in humans. One approach to the problem is genetic engineering of the fungal host to permit production of complex and hybrid N-glycans. UDP-GlcNAc:3-d-mannoside -1,2-N-acetylglucosaminyltransferase I (GnT I) is essential for the conversion of oligomannose to hybrid and complex N-glycans in higher eukaryotic cells. Since GnT I is not produced by fungi, we have introduced into the genome ofAspergillus nidulans the gene encoding full-length rabbit GnT I and demonstrated the expression of GnT I enzyme activity at levels appreciably higher than occurs in most mammalian tissues. All the GnT I activity in theAspergillus transformants remains intracellular suggesting that the rabbit trans-membrane sequence may be capable of targeting GnT I to the fungal Golgi apparatus.Abbreviations CM
complete medium
- Gal-T
UDP-Gal:GlcNAc -1,4-galactosyltransferase (EC 2.4.1.38/90)
- GnT I
UDP-GlcNAc:3-d-mannoside -1,2-N-acetylglucosaminyltransferase I (EC 2.4.1.101)
- HPLC
high performance liquid chromatography
- M3-octyl
Man1-6[Man1-3]Man-octyl
- PAGE
polyacrylamide gel electrophoresis
- MES
2-(N-morpholino)ethane sulfonate
- PCR
polymerase chain reaction
- PEG
polyethylene glycol
- PMSF
phenyl methyl sulfonyl fluoride
- SDS
sodium dodecyl sulfate
- SSC (1×)
0.15m NaCl/0.015m sodium citrate (pH 7.0)
- STC
1.2m sorbitol, 100mm Tris-HCl, pH 7.4, and 10mm CaCl2
- STET
0.1m NaCl, 10mm Tris-HCl, pH 8.0, 1mm EDTA, pH 8.0, 5% Triton-X-100
Deceased. This paper is dedicated to the memory of Lorne S. Reid. 相似文献
35.
Phosphatase Activity Toward Abnormally Phosphorylated τ: Decrease in Alzheimer Disease Brain 总被引:1,自引:0,他引:1
Cheng-Xin Gong Sadia Shaikh Jian-Zhi Wang Tanweer Zaidi Inge Grundke-Iqbal Khalid Iqbal 《Journal of neurochemistry》1995,65(2):732-738
Abstract: Microtubule-associated protein τ is abnormally hyperphosphorylated and aggregated in affected neurons of Alzheimer disease brain. This hyperphosphorylated τ can be dephosphorylated at some of the abnormal phosphorylated sites by purified protein phosphatase-1, 2A, and 2B in vitro. In the present study, we have developed an assay to measure protein phosphatase activity toward τ-1 sites (Ser199 /Ser202 ) using the hyperphosphorylated τ isolated from Alzheimer disease brain as substrate. Using this assay, we have identified that in normal brain, protein phosphatase-2A and 2B and, to a lesser extent, 1 are involved in the dephosphorylation of τ. The K m values of dephosphorylation of the hyperphosphorylated τ by protein phosphatase-2A and 2B are similar. The τ phosphatase activity is decreased by ∼30% in brain of Alzheimer disease patients compared with those of age-matched controls. These findings suggest that a defect of protein phosphatase could be the cause of the abnormal hyperphosphorylation of τ in Alzheimer disease. 相似文献
36.
Semliki Forest virus enters cells by receptor-mediated endocytosis. The acidic environment of the endosome triggers a membrane fusion reaction that is mediated by the E1 glycoprotein. During fusion, E1 rearranges from an E1/E2 heterodimer to a highly stable, membrane-inserted E1 homotrimer (E1HT). In this study, we analyzed E1HT by a combination of electron cryomicroscopy, electron crystallography of negatively stained 2D crystals, and fitting of the available X-ray structure of the monomeric E1 ectodomain into the resulting 3D reconstruction. The visualized E1HT reveals that the ectodomain has reoriented vertically and inserted the distal tip of domain II into the lipid bilayer. Our data allow the visualization of a viral fusion protein inserted in its target membrane and demonstrate that insertion is a cooperative process, resulting in rings composed of five to six homotrimers. 相似文献
37.
38.
39.
Yosepha Shahak Christof Klughammer Ulrich Schreiber Etana Padan Inge Herrman Günter Hauska 《Photosynthesis research》1994,39(2):175-181
The reduction by sulfide of exogenous ubiquinone is compared to the reduction of cytochromes in chromatophores of Rhodobacter capsulatus. From titrations with sulfide values for Vmax of 300 and 10 moles reduced/mg bacteriochlorophyll a·h, and for Km of 5 and 3 M were estimated, for decyl-ubiquinone-and cytochrome c-reduction, respectively. Both reactions are sensitive to KCN, as has been found for sulfide-quinone reductase (SQR) in Oscillatoria limnetica, which is a flavoprotein. Effects of inhibitors interfering with quinone binding sites suggest that at least part of the electron transport from sulfide in R. capsulatus employs the cytochrome bc
1-complex via the ubiquinone pool.Abbreviations BChl a
bacteriochlorophyll a
- DAD
diaminodurene
- decyl-UQ
decyl-ubiquinone
- LED
light emitting diode
- NQNO
2-n-nonyl-4-hydroxyquinoline-N-oxide
- PQ-1
plastoquinone 1
- SQR
sulfide-quinone reductase (E.C. 1.8.5.'.)
- UQ
ubiquinone 10
- Qc
the quinone reduction site on the cytochrome b
6
f/bc
1, complex (also termed Qi or Qr or Qn)
- Qs
the quinone reduction site on SQR
- Qz
quinol oxidation site on the b
6
f/bc
1, complex (also termed Qo or Qp) 相似文献
40.
Vaccination with p53-peptide–pulsed dendritic cells,of patients with advanced breast cancer: report from a phase I study 总被引:3,自引:0,他引:3
Svane IM Pedersen AE Johnsen HE Nielsen D Kamby C Gaarsdal E Nikolajsen K Buus S Claesson MH 《Cancer immunology, immunotherapy : CII》2004,53(7):633-641
Peptides derived from over-expressed p53 protein are presented by class I MHC molecules and may act as tumour-associated epitopes. Due to the diversity of p53 mutations, immunogenic peptides representing wild-type sequences are preferable as a basis for a broad-spectrum p53-targeting cancer vaccine. Our preclinical studies have shown that wild-type p53-derived HLA-A2–binding peptides are able to activate human T cells and that the generated effector T cells are cytotoxic to human HLA-A2+, p53+ tumour cells. In this phase I pilot study, the toxicity and efficacy of autologous dendritic cells (DCs) loaded with a cocktail of three wild-type and three modified p53 peptides are being analysed in six HLA-A2+ patients with progressive advanced breast cancer. Vaccinations were well tolerated and no toxicity was observed. Disease stabilisation was seen in two of six patients, one patient had a transient regression of a single lymph node and one had a mixed response. ELISpot analyses showed that the p53-peptide–loaded DCs were able to induce specific T-cell responses against modified and unmodified p53 peptides in three patients, including two of the patients with a possible clinical benefit from the treatment. In conclusion, the strategy for p53-DC vaccination seems safe and without toxicity. Furthermore, indications of both immunologic and clinical effect were found in heavily pretreated patients with advanced breast cancer. An independent clinical effect of repeated administration of DCs and IL-2 can not of course be excluded; further studies are necessary to answer these questions. 相似文献