全文获取类型
收费全文 | 289篇 |
免费 | 29篇 |
专业分类
318篇 |
出版年
2022年 | 3篇 |
2021年 | 9篇 |
2019年 | 3篇 |
2016年 | 2篇 |
2015年 | 6篇 |
2014年 | 7篇 |
2013年 | 9篇 |
2012年 | 21篇 |
2011年 | 13篇 |
2010年 | 19篇 |
2009年 | 18篇 |
2008年 | 19篇 |
2007年 | 12篇 |
2006年 | 8篇 |
2005年 | 14篇 |
2004年 | 9篇 |
2003年 | 8篇 |
2002年 | 4篇 |
2001年 | 6篇 |
2000年 | 5篇 |
1999年 | 10篇 |
1998年 | 5篇 |
1997年 | 2篇 |
1996年 | 6篇 |
1995年 | 2篇 |
1994年 | 3篇 |
1993年 | 2篇 |
1992年 | 6篇 |
1991年 | 10篇 |
1990年 | 2篇 |
1989年 | 4篇 |
1988年 | 4篇 |
1987年 | 6篇 |
1986年 | 3篇 |
1985年 | 5篇 |
1984年 | 5篇 |
1982年 | 3篇 |
1981年 | 4篇 |
1980年 | 4篇 |
1979年 | 4篇 |
1978年 | 4篇 |
1977年 | 7篇 |
1975年 | 3篇 |
1974年 | 5篇 |
1973年 | 5篇 |
1971年 | 1篇 |
1966年 | 1篇 |
1926年 | 1篇 |
1923年 | 1篇 |
1922年 | 1篇 |
排序方式: 共有318条查询结果,搜索用时 0 毫秒
1.
2.
A poxvirus-encoded uracil DNA glycosylase is essential for virus viability. 总被引:11,自引:7,他引:4 下载免费PDF全文
Infection of cultured mammalian cells with the Leporipoxvirus Shope fibroma virus (SFV) causes the induction of a novel uracil DNA glycosylase activity in the cytoplasms of the infected cells. The induction of this activity, early in infection, correlates with the early expression of the SFV BamHI D6R open reading frame which possesses significant protein sequence similarity to eukaryotic and prokaryotic uracil DNA glycosylases. The SFV BamHI D6R open reading frame and the homologous HindIII D4R open reading frame from the Orthopoxvirus vaccinia virus were cloned under the regulation of a phage T7 promoter and expressed in Escherichia coli as insoluble high-molecular-weight aggregates. During electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, the E. coli-expressed proteins migrate with an apparent molecular mass of 25 kDa. The insoluble protein aggregate generated by expression in E. coli was solubilized in urea and, following a subsequent refolding step, displayed the ability to excise uracil residues from double-stranded plasmid DNA substrates, with the subsequent formation of apyrimidinic sites. The viral enzyme, like all other characterized uracil DNA glycosylases, is active in the presence of high concentrations of EDTA, is substrate inhibited by uracil, and does not display any endonuclease activity. Attempts to inactivate the HindIII D4R gene of vaccinia virus by targeted insertion of a dominant xanthine-guanine phosphoribosyltransferase selection marker or direct insertion of a frame-shifted oligonucleotide were uniformly unsuccessful demonstrating that, unlike the uracil DNA glycosylase described for herpesviruses, the poxvirus enzyme is essential for virus viability. 相似文献
3.
Assembly of vaccinia virus: incorporation of p14 and p32 into the membrane of the intracellular mature virus. 总被引:8,自引:7,他引:1 下载免费PDF全文
B Sodeik S Cudmore M Ericsson M Esteban E G Niles G Griffiths 《Journal of virology》1995,69(6):3560-3574
The cytoplasmic assembly of vaccinia virus begins with the transformation of a two-membraned cisterna derived from the intermediate compartment between the endoplasmic reticulum and the Golgi complex. This cisterna develops into a viral crescent which eventually forms a spherical immature virus (IV) that matures into the intracellular mature virus (IMV). Using immunoelectron microscopy, we determined the subcellular localization of p32 and p14, two membrane-associated proteins of vaccinia virus. p32 was associated with vaccinia virus membranes at all stages of virion assembly, starting with the viral crescents, as well as with the membranes which accumulated during the inhibition of assembly by rifampin. There was also low but significant labelling of membranes of some cellular compartments, especially those in the vicinity of the Golgi complex. In contrast, anti-p14 labelled neither the crescents nor the IV but gave strong labelling of an intermediate form between IV and IMV and was then associated with all later viral forms. This protein was also not significantly detected on identifiable cellular membranes. Both p32 and p14 were abundantly expressed on the surface of intact IMV. Our data are consistent with a model whereby p32 would become inserted into cellular membranes before being incorporated into the crescents whereas p14 would be posttranslationally associated with the viral outer membrane at a specific later stage of the viral life cycle. 相似文献
4.
5.
6.
7.
Computer detection of the rapid diffusion of fluorescent membrane fusion markers in images observed with video microscopy. 下载免费PDF全文
We have developed an algorithm for automated detection of the dynamic pattern characterizing flashes of fluorescence in video images of membrane fusion. The algorithm detects the spatially localized, transient increases and decreases in brightness that result from the dequenching of fluorescent dye in phospholipid vesicles or lipid-enveloped virions fusing with a planar membrane. The flash is identified in video images by its nonzero time derivative and the symmetry of its spatial profile. Differentiation is implemented by forward and backward subtractions of video frames. The algorithm groups spatially connected pixels brighter than a user-specified threshold into distinct objects in forward- and backward-differentiated images. Objects are classified as either flashes or noise particles by comparing the symmetries of matched forward and backward difference profiles and then by tracking each profile in successive difference images. The number of flashes identified depends on the brightness threshold, the size of the convolution kernel used to filter the image, and the time difference between the subtracted video frames. When these parameters are changed so that the algorithm identifies an increasing percentage of the flashes recognized by eye, an increasing number of noise objects are mistakenly identified as flashes. These mistaken flashes can be eliminated by a human observer. The algorithm considerably shortens the time needed to analyze video data. Tested extensively with phospholipid vesicle and virion fusion with planar membranes, our implementation of the algorithm accurately determined the rate of fusion of influenza virions labeled with the lipophilic dye octadecylrhodamine (R18). 相似文献
8.
9.
Topological constraints in nucleic acid hybridization kinetics 总被引:2,自引:0,他引:2
Bois JS Venkataraman S Choi HM Spakowitz AJ Wang ZG Pierce NA 《Nucleic acids research》2005,33(13):4090-4095
A theoretical examination of kinetic mechanisms for forming knots and links in nucleic acid structures suggests that molecules involving base pairs between loops are likely to become topologically trapped in persistent frustrated states through the mechanism of ‘helix-driven wrapping’. Augmentation of the state space to include both secondary structure and topology in describing the free energy landscape illustrates the potential for topological effects to influence the kinetics and function of nucleic acid strands. An experimental study of metastable complementary ‘kissing hairpins’ demonstrates that the topological constraint of zero linking number between the loops effectively prevents conversion to the minimum free energy helical state. Introduction of short catalyst strands that break the topological constraint causes rapid conversion to full duplex. 相似文献
10.
Mona N. Högberg Róbert Blaško Lisbet Holm Bach Niles J. Hasselquist Gustaf Egnell Torgny Näsholm Peter Högberg 《Plant and Soil》2014,381(1-2):45-60