全文获取类型
收费全文 | 34260篇 |
免费 | 2987篇 |
国内免费 | 3820篇 |
专业分类
41067篇 |
出版年
2024年 | 109篇 |
2023年 | 500篇 |
2022年 | 1112篇 |
2021年 | 1791篇 |
2020年 | 1243篇 |
2019年 | 1598篇 |
2018年 | 1511篇 |
2017年 | 1171篇 |
2016年 | 1570篇 |
2015年 | 2150篇 |
2014年 | 2602篇 |
2013年 | 2795篇 |
2012年 | 3171篇 |
2011年 | 2934篇 |
2010年 | 1862篇 |
2009年 | 1692篇 |
2008年 | 1935篇 |
2007年 | 1716篇 |
2006年 | 1503篇 |
2005年 | 1294篇 |
2004年 | 1043篇 |
2003年 | 943篇 |
2002年 | 807篇 |
2001年 | 548篇 |
2000年 | 507篇 |
1999年 | 484篇 |
1998年 | 306篇 |
1997年 | 266篇 |
1996年 | 250篇 |
1995年 | 218篇 |
1994年 | 211篇 |
1993年 | 147篇 |
1992年 | 205篇 |
1991年 | 154篇 |
1990年 | 162篇 |
1989年 | 110篇 |
1988年 | 73篇 |
1987年 | 62篇 |
1986年 | 64篇 |
1985年 | 51篇 |
1984年 | 33篇 |
1983年 | 35篇 |
1982年 | 28篇 |
1981年 | 20篇 |
1980年 | 10篇 |
1979年 | 19篇 |
1978年 | 7篇 |
1977年 | 9篇 |
1976年 | 9篇 |
1975年 | 9篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
31.
32.
The availability of DNA structural probes that can be applied to living cells is essential for the analysis of biological functions of unusual DNA structures adopted in vivo. We have developed a chemical probe assay to detect and quantitate left-handed Z-DNA structures in recombinant plasmids in growing E. coli cells. Potassium permanganate selectively reacts with B-Z or Z-Z junction regions in supercoiled plasmids harbored in the cells. Restriction enzyme recognition sites located at these junctions are not cleaved by the corresponding endonuclease after modification with KMnO4. This inhibition of cleavage allows the determination of the relative amounts of B- and Z-forms of the cloned inserts inside the cell. We have successfully applied this method to monitor the extent of Z-DNA formation in E. coli as a function of the growth phase and mutated topoisomerase or gyrase activities. The assay can in principle be used for any unusual DNA structure that contains a restriction recognition site inside or near the structural alteration. It can be a useful tool to analyze in vivo correlations between DNA structure and gene regulatory events. 相似文献
33.
34.
Ailiang Jiang 《International journal of biometeorology》1988,32(4):280-282
According to the author's and his collaborators' investigations, the climate influences the growth of rubber trees (Hevea brasiliensis) in Xishuangbanna, the southern part of Yunnan Province, China, in at least four aspects: (1) The yield of latex per tapping and the final yield of dry rubber per tree per year or per unit area per year; (2) the growth rate, as expressed by increment of girth in cm; (3) the survival during the over-wintering period; (4) the initiation or suppression of certain diseases; In this paper the author would like to describe the influence of climatic elements on yield of latex and on survival during the over-wintering period. As for the other two aspects, only general comments are given. 相似文献
35.
Kinetic measurements of fusion of phosphatidylserine-containing vesicles by electron microscopy and fluorometry 总被引:1,自引:0,他引:1
Large unilamellar vesicles (REV) containing phosphatidylserine and phosphatidylethanolamine at a ratio of 1:3 were induced to fuse by adding calcium (4 mM). The kinetics of fusion was monitored by fluorometry using terbium or dipicolinic acid-containing vesicles. The morphology and the states of vesicle aggregation and fusion were examined at approx. 2, 30, 60, 150 and 900 s after calcium addition, by rapid quenching and freeze-fracture electron microscopy. The size and the state of aggregation of vesicles are quantitated from 4000 randomly selected vesicles. The aggregation and fusion kinetics as assayed by fluorescence volume mixing is very well simulated and predicted by the mass action model. The model essentially predicts the time course of the distribution of the aggregates and the increase in size of fused particles as measured by electron microscopy, although in some cases the predicted fusion rate exceeds that by morphometric measurement. No morphological features can be defined as fusion intermediates, although bead-like and rim-like materials may be attributed to the remnants of broken diaphragms between fusion partners. 相似文献
36.
37.
Summary Factors affecting the polyethylene glycol (PEG)-induced membrane fusion were examined. Human erythrocyte membrane ghosts, cytoskeleton-free vesicles budded from erythrocytes, mechanically disrupted erythrocyte vesicles, and recombinant vesicles from glycophorin and egg phosphatidylcholine were used as models. Fusion was monitored by darkfield light microscopy and by freeze-fracture electron microscopy. Osmotic swelling was found necessary for fusion between membrane ghosts following PEG treatment. The sample with the highest fusion percentage was sealed ghosts incubated in hypotonic media after at least 5 min of treatment in <25% PEG. At similar osmolarity, glycerol, dextran and PEG produced progressively more pronounced intramembranous particle (IMP) patching, correlating with their increasing fusion percentages. The patching of IMP preceded cell-cell contact, and occurred without direct PEG-protein interaction. The presence of cytoskeletal elements in small vesicles had no significant effect on fusion, nor on the aggregation of intramembranous particle (IMP) upon PEG treatment. Disrupting the membrane by lysolecithin, dimethylsulfoxide, retinol or mild sonication resulted in the fragmentation of ghosts without an increase in fusion percentage. The purity of the commercial PEG used had no apparent effect on fusion. We concluded that the key steps in PEG-induced fusion of cell membrane are the creation of IMP-free zones, and the osmotic swelling of cells after the formation of bilayer contacts during the PEG treatment. Cell cytoskeleton affects PEG-induced fusion only to the extent of affecting IMP patching. 相似文献
38.
39.
40.
Proteases from human immunodeficiency virus and avian myeloblastosis virus show distinct specificities in hydrolysis of multidomain protein substrates. 下载免费PDF全文
A G Tomasselli J O Hui T K Sawyer D J Staples C A Bannow I M Reardon V K Chaudhary C M Fryling I Pastan D J Fitzgerald et al. 《Journal of virology》1990,64(7):3157-3161
The virally encoded proteases from human immunodeficiency virus (HIV) and avian myeloblastosis virus (AMV) have been compared relative to their ability to hydrolyze a variant of the three-domain Pseudomonas exotoxin, PE66. This exotoxin derivative, missing domain I and referred to as LysPE40, is made up of a 13-kilodalton NH2-terminal translocation domain II connected by a segment of 40 amino acids to enzyme domain III of the toxin, a 23-kilodalton ADP-ribosyltransferase. HIV protease hydrolyzes two peptide bonds in LysPE40, a Leu-Leu bond in the interdomain region and a Leu-Ala bond in a nonstructured region three residues in from the NH2-terminus. Neither of these sites is cleaved by the AMV enzyme; hydrolysis occurs, instead, at an Asp-Val bond in another part of the interdomain segment and at a Leu-Thr bond in the NH2-terminal region of domain II. Synthetic peptides corresponding to these cleavage sites are hydrolyzed by the individual proteases with the same specificity displayed toward the protein substrate. Peptide substrates for one protease are neither substrates nor competitive inhibitors for the other. A potent inhibitor of HIV type 1 protease was more than 3 orders of magnitude less active toward the AMV enzyme. These results suggest that although the crystallographic models of Rous sarcoma virus protease (an enzyme nearly identical to the AMV enzyme) and HIV type 1 protease show a high degree of similarity, there exist structural differences between these retroviral proteases that are clearly reflected by their kinetic properties. 相似文献