全文获取类型
收费全文 | 2834篇 |
免费 | 243篇 |
国内免费 | 5篇 |
专业分类
3082篇 |
出版年
2024年 | 2篇 |
2023年 | 11篇 |
2022年 | 38篇 |
2021年 | 87篇 |
2020年 | 46篇 |
2019年 | 67篇 |
2018年 | 79篇 |
2017年 | 89篇 |
2016年 | 93篇 |
2015年 | 157篇 |
2014年 | 158篇 |
2013年 | 220篇 |
2012年 | 202篇 |
2011年 | 215篇 |
2010年 | 137篇 |
2009年 | 131篇 |
2008年 | 182篇 |
2007年 | 221篇 |
2006年 | 187篇 |
2005年 | 170篇 |
2004年 | 154篇 |
2003年 | 121篇 |
2002年 | 136篇 |
2001年 | 18篇 |
2000年 | 10篇 |
1999年 | 17篇 |
1998年 | 25篇 |
1997年 | 18篇 |
1996年 | 10篇 |
1995年 | 6篇 |
1994年 | 9篇 |
1993年 | 13篇 |
1992年 | 9篇 |
1990年 | 6篇 |
1989年 | 4篇 |
1988年 | 2篇 |
1987年 | 2篇 |
1986年 | 4篇 |
1983年 | 4篇 |
1981年 | 2篇 |
1980年 | 2篇 |
1979年 | 2篇 |
1978年 | 2篇 |
1977年 | 3篇 |
1972年 | 1篇 |
1971年 | 1篇 |
1970年 | 1篇 |
1969年 | 1篇 |
1967年 | 1篇 |
1964年 | 1篇 |
排序方式: 共有3082条查询结果,搜索用时 15 毫秒
1.
Formaldehyde fixation of cells is routinely used to study DNA-protein interactions in vivo. In these studies, DNA is often analyzed using a polymerase chain reaction technique. Although it is known that formaldehyde can damage DNA, no studies have been performed so far to compare the efficiency of DNA amplification between normal and fixed cells. Here we show that formaldehyde fixation results in a 15% to 20% reduction in the ability to amplify cellular DNA. The loss of amplifiability is independent of the length of the amplification region and the degree to which DNA is compacted on packaging into chromatin. 相似文献
2.
3.
R P Morozova V A Kokunin I A Nikolenko N V Kanivets M V Stefanov 《Ukrainski? biokhimicheski? zhurnal》1988,60(4):18-24
DEAE-cellulose chromatography and SDS-PAAG electrophoresis were used to study proteins from endoplasmic reticulum of smooth and rough enterocyte membranes in the vitamin D-replete and deficient rat. The membrane-bound vitamin-D-dependent protein with a molecular weight of 21-25 kDa is shown to consist of the endoplasmic reticulum of smooth membranes. The intensive binding of calcium by these membranes is established. 相似文献
4.
Deletion analysis of the SUP35 gene of the yeast Saccharomyces cerevisiae reveals two non-overlapping functional regions in the encoded protein 总被引:13,自引:0,他引:13
Michael D. Ter-Avanesyan Vitaly V. Kushnirov Adilya R. Dagkesamanskaya Svetlana A. Didichenko Yury O. Chernoff Sergey G. Inge-Vechtomov Vladimir N. Smirnov 《Molecular microbiology》1993,7(5):683-692
SUP35is an omnipotent suppressor gene of Saccharomyces cerevisiae coding for a protein consisting of a C-terminal part similar to the elongation factor EF-1α and a unique N-terminal sequence of 253 amino acids. Twelve truncated versions of the SUP35 gene were generated by the deletion of fragments internal to the coding sequence. Functional studies of these deletion mutants showed that: (i) only the EF-1α-like C-terminal part of the Sup35 protein is essential for the cell viability; (ii) overexpression of either the N-terminal part of the Sup35 protein or the full-length Sup35 protein decreases translational fidelity, resulting in omnipotent suppression and reduced growth of [psi+] strains; (iii) expression of the C-terminal part of the Sup35 protein generates an antisuppressor phenotype; and (iv) both the N- or C-terminal segments of the Sup35 protein can bind to 80S ribosomes. Thus, the data obtained define two domains within the Sup35 protein which are responsible for different functions. 相似文献
5.
Bernhard H. J. Juurlink Sergey Fedoroff 《In vitro cellular & developmental biology. Plant》1979,15(2):86-94
Summary Whole mouse embryos were grown in vitro from Theiler stage 12 (1 to 7 somites) to Theiler stages 15 and 16 (25 to 35 somites).
This procedure gives experimental access to precisely staged embryos during the early period of neurogenesis. To follow the
further development of neurons in vitro, fragments of spinal primordia were set up from these cultured embryos. In such cultures,
the proliferation of precursor cells, the formation of postmitotic cells and, finally, the cytodifferentiation of neurons
were observed.
A preliminary account of this work was given at the Tissue Culture Association Meeting in 1977, and the Canadian Federation
of Biological Societies Meeting in 1977 (1,2).
This work was supported by Grant MT 4235 from the Medical Research Council of Canada. 相似文献
6.
Sergey Ivanov Maria J. Harrison 《The Plant journal : for cell and molecular biology》2014,80(6):1151-1163
Medicago truncatula is widely used for analyses of arbuscular mycorrhizal (AM) symbiosis and nodulation. To complement the genetic and genomic resources that exist for this species, we generated fluorescent protein fusions that label the nucleus, endoplasmic reticulum, Golgi apparatus, trans‐Golgi network, plasma membrane, apoplast, late endosome/multivesicular bodies (MVB), transitory late endosome/ tonoplast, tonoplast, plastids, mitochondria, peroxisomes, autophagosomes, plasmodesmata, actin, microtubules, periarbuscular membrane (PAM) and periarbuscular apoplastic space (PAS) and expressed them from the constitutive AtUBQ10 promoter and the AM symbiosis‐specific MtBCP1 promoter. All marker constructs showed the expected expression patterns and sub‐cellular locations in M. truncatula root cells. As a demonstration of their utility, we used several markers to investigate AM symbiosis where root cells undergo major cellular alterations to accommodate their fungal endosymbiont. We demonstrate that changes in the position and size of the nuclei occur prior to hyphal entry into the cortical cells and do not require DELLA signaling. Changes in the cytoskeleton, tonoplast and plastids also occur in the colonized cells and in contrast to previous studies, we show that stromulated plastids are abundant in cells with developing and mature arbuscules, while lens‐shaped plastids occur in cells with degenerating arbuscules. Arbuscule development and secretion of the PAM creates a periarbuscular apoplastic compartment which has been assumed to be continuous with apoplast of the cell. However, fluorescent markers secreted to the periarbuscular apoplast challenge this assumption. This marker resource will facilitate cell biology studies of AM symbiosis, as well as other aspects of legume biology. 相似文献
7.
Development of microbial biofilms and the recruitment of propagules on the surfaces of man-made structures in the marine environment cause serious problems for the navies and for marine industries around the world. Current antifouling technology is based on the application of toxic substances that can be harmful to the natural environment. For this reason and the global ban of tributyl tin (TBT), there is a need for the development of "environmentally-friendly" antifoulants. Marine microbes are promising potential sources of non-toxic or less-toxic antifouling compounds as they can produce substances that inhibit not only the attachment and/or growth of microorganisms but also the settlement of invertebrate larvae and macroalgal spores. However, so far only few antilarval settlement compounds have been isolated and identified from bacteria. In this review knowledge about antifouling compounds produced by marine bacteria and diatoms are summarised and evaluated and future research directions are highlighted. 相似文献
8.
Sergey N Arkhipov Maxim Berezovski Julia Jitkova Sergey N Krylov 《Cytometry. Part A》2005,63(1):41-47
BACKGROUND: Chemical cytometry is an emerging technology that analyzes chemical contents of single cells by means of capillary electrophoresis or capillary chromatography. It has a potential to become an indispensable tool in analyses of heterogeneous cell populations such as those in tumors. Ras oncogenes are found in 30% of human cancers. To become fully functional products, oncogenic Ras proteins require at least three posttranslational modifications: farnesylation, endoproteolysis, and carboxyl-methylation. Therefore, enzymes that catalyze the three reactions, farnesyltransferase (FTase), endoprotease (EPase), and methyltransferase (MTase), are considered highly attractive therapeutic targets. In this work, we used chemical cytometry to study the metabolism of a pentapeptide substrate that can mimic Ras proteins with respect to their posttranslational modifications in solution. METHODS: Mouse mammary gland tumor cells (4T1) and mouse embryo fibroblasts (NIH3T3) were incubated with a fluorescently labeled pentapeptide substrate, 2',7'-difluorofluorescein-5-carboxyl-Gly-Cys-Val-Ilu-Ala. Cells were washed from the substrate and resuspended in phosphate buffered saline. Uptake of the substrate by the cells was monitored by laser scanning confocal microscopy. Single cells were injected into the capillary, lysed, and subjected to capillary electrophoresis. Fluorescent metabolic products were detected by laser-induced fluorescence and compared with products obtained by the conversion of the substrate by FTase, EPase, and MTase in solution. Co-sampling of single cells with the in-vitro products was used for such comparison. RESULTS: Confocal microscopy data showed that the substrate permeated the plasma membrane and clustered in the cytoplasm. Further capillary electrophoresis and chemical cytometry analyses showed that the substrate was converted into three fluorescently labeled products, two of which were secreted in the culture medium and one remained in the cells. The intracellular product was present at approximately 100,000 molecules per cell. The three metabolic products of the substrate were found to be different from the products of its processing by FTase, EPase, and MTase in solution. CONCLUSIONS: This is the first report of chemical cytometry in the context of Ras-signaling studies. The chemical cytometry method used in this work will find applications in the development of suitable peptide substrates for monitoring enzyme activities in single cells. 相似文献
9.
Tatiana S. Cherkesova Tatiana Y. Hargrove M. Cristina Vanrell Igor Ges Sergey A. Usanov Patricia S. Romano Galina I. Lepesheva 《FEBS letters》2014
CYP51 (sterol 14α-demethylase) is an efficient target for clinical and agricultural antifungals and an emerging target for treatment of Chagas disease, the infection that is caused by multiple strains of a protozoan pathogen Trypanosoma cruzi. Here, we analyze CYP51A from the Y strain T. cruzi. In this protein, proline 355, a residue highly conserved across the CYP51 family, is replaced with serine. The purified enzyme retains its catalytic activity, yet has been found less susceptible to inhibition. These biochemical data are consistent with cellular experiments, both in insect and human stages of the pathogen. Comparative structural analysis of CYP51 complexes with VNI and two derivatives suggests that broad-spectrum CYP51 inhibitors are likely to be preferable as antichagasic drug candidates. 相似文献