全文获取类型
收费全文 | 604篇 |
免费 | 50篇 |
国内免费 | 32篇 |
出版年
2022年 | 4篇 |
2021年 | 6篇 |
2020年 | 8篇 |
2019年 | 4篇 |
2018年 | 14篇 |
2017年 | 8篇 |
2016年 | 12篇 |
2015年 | 18篇 |
2014年 | 22篇 |
2013年 | 29篇 |
2012年 | 33篇 |
2011年 | 46篇 |
2010年 | 32篇 |
2009年 | 24篇 |
2008年 | 29篇 |
2007年 | 23篇 |
2006年 | 36篇 |
2005年 | 22篇 |
2004年 | 14篇 |
2003年 | 20篇 |
2002年 | 18篇 |
2001年 | 16篇 |
2000年 | 10篇 |
1999年 | 8篇 |
1998年 | 14篇 |
1997年 | 9篇 |
1996年 | 7篇 |
1995年 | 6篇 |
1994年 | 7篇 |
1992年 | 13篇 |
1991年 | 6篇 |
1990年 | 10篇 |
1989年 | 8篇 |
1988年 | 13篇 |
1987年 | 14篇 |
1986年 | 5篇 |
1985年 | 8篇 |
1984年 | 7篇 |
1983年 | 7篇 |
1982年 | 5篇 |
1981年 | 7篇 |
1980年 | 6篇 |
1977年 | 9篇 |
1975年 | 6篇 |
1973年 | 8篇 |
1971年 | 4篇 |
1970年 | 3篇 |
1958年 | 3篇 |
1955年 | 3篇 |
1949年 | 5篇 |
排序方式: 共有686条查询结果,搜索用时 390 毫秒
41.
42.
Escherichia coli polymerase 1 (Pol 1) and Thermus aquaticus Taq polymerase are homologous Type I DNA polymerases, each comprised of a polymerase domain, a proofreading domain (inactive in Taq), and a 5' nuclease domain. "Klenow" and "Klentaq" are the large fragments of Pol 1 and Taq and are functional polymerases lacking the 5' nuclease domain. In the available crystal structures of full-length Taq, the 5' nuclease domain is positioned in two different orientations: in one structure, it is extended out into solution, whereas in the other, it is folded up against the polymerase domain in a more compact structure. Analytical ultracentrifugation experiments report s20,w values of 5.05 for Taq, 4.1 for Klentaq, 5.3 for E. coli Pol 1, and 4.6 for Klenow. Measured partial specific volumes are all quite similar, indicating no significant differences in packing density between the mesophilic and thermophilic proteins. Small angle x-ray scattering studies report radii of gyration of 38.3 A for Taq, 30.7 A for Klentaq, and 30.5 A for Klenow. The hydrodynamic and x-ray scattering properties of the polymerases were also calculated directly from the different crystal structures using the programs HYDROPRO (Garcia De La Torre, J., Huertas, M. L., and Carrasco, B. (2000) Biophys J. 78, 719-730) and CRYSOL (Svergun, D. I., Barberato, C., and Koch, M. H. J. (1995) J. Appl. Crystalogr. 28, 768-773), respectively. The combined experimental and computational characterizations indicate that the full-length polymerases in solution are in a conformation where the 5' nuclease domain is extended into solution. Further, the radius of gyration, and hence the global conformation of Taq polymerase, is not altered by the binding of either matched primer template DNA or ddATP. 相似文献
43.
Comparative vaccine studies in HLA-A2.1-transgenic mice reveal a clustered organization of epitopes presented in hepatitis C virus natural infection 总被引:4,自引:0,他引:4
下载免费PDF全文
![点击此处可从《Journal of virology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Himoudi N Abraham JD Fournillier A Lone YC Joubert A Op De Beeck A Freida D Lemonnier F Kieny MP Inchauspé G 《Journal of virology》2002,76(24):12735-12746
A polyepitopic CD8(+)-T-cell response is thought to be critical for control of hepatitis C virus (HCV) infection. Using transgenic mice, we analyzed the immunogenicity and dominance of most known HLA-A2.1 epitopes presented during infection by using vaccines that carry the potential to enter clinical trials: peptides, DNA, and recombinant adenoviruses. The vaccines capacity to induce specific cytotoxic T lymphocytes and interferon gamma-producing cells revealed that immunogenic epitopes are clustered in specific antigens. For two key antigens, flanking regions were shown to greatly enhance the scope of epitope recognition, whereas a DNA-adenovirus prime-boost vaccination strategy augmented epitope immunogenicity, even that of subdominant ones. The present study reveals a clustered organization of HCV immunogenic HLA.A2.1 epitopes and strategies to modulate their dominance. 相似文献
44.
High rate of DNA loss in the Drosophila melanogaster and Drosophila virilis species groups 总被引:6,自引:3,他引:3
We recently proposed that patterns of evolution of non-LTR
retrotransposable elements can be used to study patterns of spontaneous
mutation. Transposition of non-LTR retrotransposable elements commonly
results in creation of 5' truncated, "dead-on-arrival" copies. These
inactive copies are effectively pseudogenes and, according to the neutral
theory, their molecular evolution ought to reflect rates and patterns of
spontaneous mutation. Maximum parsimony can be used to separate the
evolution of active lineages of a non-LTR element from the fate of the
"dead-on-arrival" insertions and to directly assess the relative
frequencies of different types of spontaneous mutations. We applied this
approach using a non-LTR element, Helena, in the Drosophila virilis group
and have demonstrated a surprisingly high incidence of large deletions and
the virtual absence of insertions. Based on these results, we suggested
that Drosophila in general may exhibit a high rate of spontaneous large
deletions and have hypothesized that such a high rate of DNA loss may help
to explain the puzzling dearth of bona fide pseudogenes in Drosophila. We
also speculated that variation in the rate of spontaneous deletion may
contribute to the divergence of genome size in different taxa by affecting
the amount of superfluous "junk" DNA such as, for example, pseudogenes or
long introns. In this paper, we extend our analysis to the D. melanogaster
subgroup, which last shared a common ancestor with the D. virilis group
approximately 40 MYA. In a different region of the same transposable
element, Helena, we demonstrate that inactive copies accumulate deletions
in species of the D. melanogaster subgroup at a rate very similar to that
of the D. virilis group. These results strongly suggest that the high rate
of DNA loss is a general feature of Drosophila and not a peculiar property
of a particular stretch of DNA in a particular species group.
相似文献
45.
46.
Christiaan Jacob Johan Vorster Anna Margaretha Joubert 《Cell biochemistry and function》2010,28(5):412-419
A priority in recent anti‐cancer drug development has been attaining better side‐effect profiles for potential compounds. To produce highly specific cancer therapies it is necessary to understand both the effects of the proposed compound on cancer and on normal cells comprising the rest of the human body. Thus in vitro evaluation of these compounds against non‐carcinogenic cell lines is of critical importance. One of the most recent developments in experimental anti‐cancer agents is 2‐methoxyestradiol‐bis‐sulphamate (2ME‐BM), a sulphamoylated derivative of 2‐methoxyestradiol. The aim of this study was to evaluate the in vitro effects of 2ME‐BM on cell proliferation, morphology and mechanisms of cell death in the non‐carcinogenic MCF‐12A breast epithelial cell line. The study revealed changes in proliferative capacity, morphology and cell death induction in response to 2ME‐BM exposure (24 h at 0.4 µM). Microscopy showed decreased cell density and cell death‐associated morphology (increased apoptotic characteristics), a slight increase in acidic intracellular vesicles and insignificant ultra‐structural aberrations. Mitotic indices revealed a G2M‐phase cell cycle block. This was confirmed by flow cytometry, where an increased fraction of abnormal cells and a decrease in cyclin B1 levels were observed. These results evidently demonstrate that the non‐carcinogenic MCF‐12A cell line is less susceptible when compared to 2ME‐BM‐exposed cancer cell lines previously tested. Further in vitro research into the mechanism of this potentially useful compound is warranted. Copyright © 2010 John Wiley & Sons, Ltd. 相似文献
47.
Christine Jacomet Clotilde Allavena Fleur Peyrol Bruno Pereira Laurence Morand Joubert Haleh Bagheri Laurent Cotte Rodolphe Garaffo Laurent Gerbaud Pierre Dellamonica 《PloS one》2015,10(2)
BackgroundIn the interest of cost effectiveness, switching antiretroviral brand name medications to generics is recommended in France since 2013. The study objective was to evaluate the perception of generics per se and antiretroviral generics in HIV-infected patients and their hospital physiciansConclusionsAcceptability of antiretroviral generics in this French population was mostly dictated by the patient’s and physician’s knowledge and use of generics overall. It should be improved with an efficient information of both patients and physicians. 相似文献
48.
Courtney Reichhardt Jose A. G. Ferreira Lydia-Marie Joubert Karl V. Clemons David A. Stevens Lynette Cegelski 《Eukaryotic cell》2015,14(11):1064-1072
Aspergillus fumigatus is commonly responsible for lethal fungal infections among immunosuppressed individuals. A. fumigatus forms biofilm communities that are of increasing biomedical interest due to the association of biofilms with chronic infections and their increased resistance to antifungal agents and host immune factors. Understanding the composition of microbial biofilms and the extracellular matrix is important to understanding function and, ultimately, to developing strategies to inhibit biofilm formation. We implemented a solid-state nuclear magnetic resonance (NMR) approach to define compositional parameters of the A. fumigatus extracellular matrix (ECM) when biofilms are formed in RPMI 1640 nutrient medium. Whole biofilm and isolated matrix networks were also characterized by electron microscopy, and matrix proteins were identified through protein gel analysis. The 13C NMR results defined and quantified the carbon contributions in the insoluble ECM, including carbonyls, aromatic carbons, polysaccharide carbons (anomeric and nonanomerics), aliphatics, etc. Additional 15N and 31P NMR spectra permitted more specific annotation of the carbon pools according to C-N and C-P couplings. Together these data show that the A. fumigatus ECM produced under these growth conditions contains approximately 40% protein, 43% polysaccharide, 3% aromatic-containing components, and up to 14% lipid. These fundamental chemical parameters are needed to consider the relationships between composition and function in the A. fumigatus ECM and will enable future comparisons with other organisms and with A. fumigatus grown under alternate conditions. 相似文献
49.
Pierre-Emmanuel Joubert Kenneth Stapleford Florence Guivel-Benhassine Marco Vignuzzi Olivier Schwartz Matthew L. Albert 《PLoS pathogens》2015,11(8)
Chikungunya virus (CHIKV), the causative agent of a major epidemic spanning five continents, is a positive stranded mRNA virus that replicates using the cell’s cap-dependent translation machinery. Despite viral infection inhibiting mTOR, a metabolic sensor controls cap-dependent translation, viral proteins are efficiently translated. Rapalog treatment, silencing of mtor or raptor genes, but not rictor, further enhanced CHIKV infection in culture cells. Using biochemical assays and real time imaging, we demonstrate that this effect is independent of autophagy or type I interferon production. Providing in vivo evidence for the relevance of our findings, mice treated with mTORC1 inhibitors exhibited increased lethality and showed a higher sensitivity to CHIKV. A systematic evaluation of the viral life cycle indicated that inhibition of mTORC1 has a specific positive effect on viral proteins, enhancing viral replication by increasing the translation of both structural and nonstructural proteins. Molecular analysis defined a role for phosphatidylinositol-3 kinase (PI3K) and MAP kinase-activated protein kinase (MnKs) activation, leading to the hyper-phosphorylation of eIF4E. Finally, we demonstrated that in the context of CHIKV inhibition of mTORC1, viral replication is prioritized over host translation via a similar mechanism. Our study reveals an unexpected bypass pathway by which CHIKV protein translation overcomes viral induced mTORC1 inhibition. 相似文献
50.