全文获取类型
收费全文 | 1853篇 |
免费 | 63篇 |
国内免费 | 74篇 |
专业分类
1990篇 |
出版年
2024年 | 1篇 |
2023年 | 6篇 |
2022年 | 9篇 |
2021年 | 9篇 |
2020年 | 15篇 |
2019年 | 23篇 |
2018年 | 48篇 |
2017年 | 25篇 |
2016年 | 23篇 |
2015年 | 37篇 |
2014年 | 125篇 |
2013年 | 113篇 |
2012年 | 92篇 |
2011年 | 145篇 |
2010年 | 133篇 |
2009年 | 161篇 |
2008年 | 176篇 |
2007年 | 192篇 |
2006年 | 128篇 |
2005年 | 131篇 |
2004年 | 73篇 |
2003年 | 74篇 |
2002年 | 86篇 |
2001年 | 49篇 |
2000年 | 33篇 |
1999年 | 30篇 |
1998年 | 21篇 |
1997年 | 9篇 |
1996年 | 2篇 |
1995年 | 1篇 |
1994年 | 1篇 |
1993年 | 4篇 |
1992年 | 2篇 |
1990年 | 3篇 |
1989年 | 2篇 |
1988年 | 2篇 |
1984年 | 1篇 |
1982年 | 1篇 |
1981年 | 1篇 |
1979年 | 1篇 |
1978年 | 1篇 |
1977年 | 1篇 |
排序方式: 共有1990条查询结果,搜索用时 0 毫秒
91.
Pil Joong Chung 《FEBS letters》2009,583(13):2249-271
Histone deacetylases (HDACs) are known to function in the nucleus. Here, we report on the organellar localization of three rice HDACs, OsSIR2b, OsHDAC6, and OsHDAC10. The 35S:OsSIR2b-GFP and 35S:OsHDAC10-GFP constructs were introduced into tobacco BY2 cells. Co-localization analysis of the green fluorescent protein and MitoTracker fluorescent signals in the transformed BY2 cells indicated that OsSIR2b and OsHDAC10 are localized in the mitochondria. Transgenic Arabidopsis lines harboring 35S:OsHDAC6-GFP and 35S:OsHDAC10-GFP constructs were similarly analyzed, revealing that OsHDAC6-GFP is localized exclusively in chloroplasts, whereas OsHDAC10-GFP is localized in both mitochondria and chloroplasts. The presence of OsHDAC6-GFP and OsHDAC10-GFP in chloroplasts was verified by immunodetection. 相似文献
92.
Kinoshita M Okamoto G Hirata T Shinomiya A Kobayashi T Kubo Y Hori H Kanamori A 《Molecular reproduction and development》2009,76(2):202-207
Easy oocyte detection in living specimens benefits various developmental biology and environmental toxicology studies. One of the earliest markers of sex differentiation in medaka (Oryzias latipes) is oocyte development. Within the field of toxicology, a simple detection method for induced oocyte in the testis (testis-ova) as a result of endocrine disruption is necessary. In this study we produced transgenic medaka whose oocytes were labeled with fluorescent proteins using the regulatory region of the 42Sp50 gene, an isoform of polypeptide elongation 1-alpha. Short (201 nt) 5'- and 3'-flanking regions were sufficient for reporter gene expression. GFP expression was first observed in female germ cells approximately 5 days post-hatching. In the mature ovaries, germ cells showed such intense fluorescence that the fluorescence was observed from outside the body wall. In contrast, very faint fluorescence was observed in the mature testes. Testis-ova, oocytes artificially induced in the testes, were also labeled with GFP. These findings indicate through the use of transgenic medaka, that detection of female germ cells was straightforward and this transgenic medaka model proves useful for tracking female germ cells in developmental and toxicological studies. 相似文献
93.
Anne Chevrel Agathe Urvoas Ines?Li de la Sierra-Gallay Magali Aumont-Nicaise Sandrine Moutel Michel Desmadril Franck Perez Alexis Gautreau Herman van Tilbeurgh Philippe Minard Marie Valerio-Lepiniec 《Bioscience reports》2015,35(4)
A family of artificial proteins, named αRep, based on a natural family of helical repeat was previously designed. αRep members are efficiently expressed, folded and extremely stable proteins. A large αRep library was constructed creating proteins with a randomized interaction surface. In the present study, we show that the αRep library is an efficient source of tailor-made specific proteins with direct applications in biochemistry and cell biology. From this library, we selected by phage display αRep binders with nanomolar dissociation constants against the GFP. The structures of two independent αRep binders in complex with the GFP target were solved by X-ray crystallography revealing two totally different binding modes. The affinity of the selected αReps for GFP proved sufficient for practically useful applications such as pull-down experiments. αReps are disulfide free proteins and are efficiently and functionally expressed in eukaryotic cells: GFP-specific αReps are clearly sequestrated by their cognate target protein addressed to various cell compartments. These results suggest that αRep proteins with tailor-made specificity can be selected and used in living cells to track, modulate or interfere with intracellular processes. 相似文献
94.
Yang JH Wylie-Sears J Bischoff J 《Biochemical and biophysical research communications》2008,374(3):512-516
The endothelium of the cardiac valves is unique compared the rest of the vasculature in its ability to undergo an endothelial-to-mesenchymal transformation (EMT) in vitro in response to transforming growth factor-β (TGF-β). EMT is a critical event during embryonic valve development, and both VEGF-A and Notch1 have been shown to function in this process. Here we investigate the effects of VEGF-A and Notch1 on EMT in clonal endothelial cell (EC) populations isolated from adult aortic valve leaflets. VEGF-A inhibited TGF-β-induced EMT. Endothelial growth, however, was not affected by VEGF-A or TGF-β. A positive role for Notch1 was revealed in three experiments: (1) TGF-β induced Notch1 mRNA in valve ECs, (2) a γ-secretase inhibitor of Notch1 signaling blocked EMT, and (3) overexpression of a ligand-independent form of Notch1 induced EMT. These results demonstrate, for the first time, that VEGF-A and Notch1 play opposing roles in regulating EMT in post-natal valve endothelium. 相似文献
95.
96.
S. Nomura K. Yamane T. Masuda F. Kawamura T. Mizukami H. Saito 《Bioscience, biotechnology, and biochemistry》2013,77(12):2637-2638
We constructed a reporter system to detect a superoxide-generating methyl viologen using SoxRS of Escherichia coli and GFP of Aequorea victoria. E. coli carrying this plasmid exhibited strong fluorescence when grown in the presence of a superoxide-generating reagent methyl viologen. The fluorescence intensity observed in the stationary phase culture of the transformant increased in response to the methyl viologen concentration in a range of 0.01 μM to 10 μM. 相似文献
97.
98.
Shang-Te Danny Hsu Caroline Behrens Lisa D. Cabrita Christopher M. Dobson 《Biomolecular NMR assignments》2009,3(1):67-72
We present here the backbone and side-chain NMR assignments of YFP Venus, a 238-residue protein that emits yellow fluorescence
in its native state. Venus is a variant of the green fluorescent protein (GFP), which has improved chromophore maturation
and brightness, and the photochemistry and photophysics of which are insensitive to experimental conditions, such as the pH
value and buffer content, making it a favourable biomarker. 相似文献
99.
100.
目的:建立长双歧杆菌NCC2705高效稳定的电转化系统,并以其为宿主表达绿色荧光蛋白(GFP),构建一个稳定的带报告基因的表达载体。方法:从双歧杆菌克隆载体pDG7中切取其在双歧杆菌中复制所必需的pMB1复制子插入质粒pUC19的多克隆位点区,并将gfp基因在双歧杆菌中进行表达;设计双歧杆菌电击转化体系,研究在不同电击电压条件下的电转效率。结果:首先构建了大肠杆菌-长双歧杆菌穿梭质粒pUB1和带有gfp基因的表达质粒pUB2,用电击法将之转化至双歧杆菌,当细菌生长到D600nm约为0.5时制备感受态细胞,在电容25μF、电阻200Ω、电压12.5 kV/cm的电击条件下进行完整质粒转化,可得到较高的转化率。结论:构建了以长双歧杆菌NCC2705为宿主的高效稳定的表达系统,为进一步研究益生菌的分子生物学和功能特性提供了基础。 相似文献