全文获取类型
收费全文 | 195778篇 |
免费 | 20391篇 |
国内免费 | 5238篇 |
专业分类
221407篇 |
出版年
2023年 | 1185篇 |
2022年 | 2797篇 |
2021年 | 4862篇 |
2020年 | 3174篇 |
2019年 | 3903篇 |
2018年 | 4209篇 |
2017年 | 3305篇 |
2016年 | 4613篇 |
2015年 | 6483篇 |
2014年 | 7626篇 |
2013年 | 8680篇 |
2012年 | 10106篇 |
2011年 | 9285篇 |
2010年 | 5789篇 |
2009年 | 5325篇 |
2008年 | 6349篇 |
2007年 | 5960篇 |
2006年 | 5515篇 |
2005年 | 4812篇 |
2004年 | 4316篇 |
2003年 | 4047篇 |
2002年 | 3781篇 |
2001年 | 11995篇 |
2000年 | 11841篇 |
1999年 | 9154篇 |
1998年 | 2507篇 |
1997年 | 2743篇 |
1996年 | 2515篇 |
1995年 | 2275篇 |
1994年 | 2033篇 |
1993年 | 1806篇 |
1992年 | 5809篇 |
1991年 | 5360篇 |
1990年 | 4703篇 |
1989年 | 4527篇 |
1988年 | 4024篇 |
1987年 | 3453篇 |
1986年 | 3045篇 |
1985年 | 2931篇 |
1984年 | 2143篇 |
1983年 | 1887篇 |
1982年 | 1322篇 |
1979年 | 1837篇 |
1978年 | 1409篇 |
1977年 | 1246篇 |
1976年 | 1080篇 |
1975年 | 1182篇 |
1974年 | 1218篇 |
1973年 | 1221篇 |
1972年 | 1082篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
21.
Minjuan Shen Mingli Lin Mengqi Zhu Wenxin Zhang Danyang Lu Huanhuan Liu Jingjing Deng Kehua Que Xu Zhang 《Biochimica et Biophysica Acta (BBA)/General Subjects》2019,1863(1):167-181
Since their discovery, matrix vesicles (MVs) containing minerals have received considerable attention for their role in the mineralization of bone, dentin and calcified cartilage. Additionally, MVs' association with collagen fibrils, which serve as the scaffold for calcification in the organic matrix, has been repeatedly highlighted. The primary purpose of the present study was to establish a MVs–mimicking model (PEG-S-ACP/micelle) in vitro for studying the exact mechanism of MVs-mediated extra/intra fibrillar mineralization of collagen in vivo. In this study, high-concentration serine was used to stabilize the amorphous calcium phosphate (S-ACP), which was subsequently mixed with polyethylene glycol (PEG) to form PEG-S-ACP nanoparticles. The nanoparticles were loaded in the polysorbate 80 micelle through a micelle self-assembly process in an aqueous environment. This MVs–mimicking model is referred to as the PEG-S-ACP/micelle model. By adjusting the pH and surface tension of the PEG-S-ACP/micelle, two forms of minerals (crystalline mineral nodules and ACP nanoparticles) were released to achieve the extrafibrillar and intrafibrillar mineralization, respectively. This in vitro mineralization process reproduced the mineral nodules mediating in vivo extrafibrillar mineralization and provided key insights into a possible mechanism of biomineralization by which in vivo intrafibrillar mineralization could be induced by ACP nanoparticles released from MVs. Also, the PEG-S-ACP/micelle model provides a promising methodology to prepare mineralized collagen scaffolds for repairing bone defects in bone tissue engineering. 相似文献
22.
23.
24.
To define catalytically essential residues of bacteriophage T7 RNA polymerase, we have generated five mutants of the polymerase, D537N, K631M, Y639F, H811Q and D812N, by site-directed mutagenesis and purified them to homogeneity. The choice of specific amino acids for mutagenesis was based upon photoaffinity-labeling studies with 8-azido-ATP and homology comparisons with the Klenow fragment and other DNA/RNA polymerases. Secondary structural analysis by circular dichroism indicates that the protein folding is intact in these mutants. The mutants D537N and D812N are totally inactive. The mutant K631M has 1% activity, confined to short oligonucleotide synthesis. The mutant H811Q has 25% activity for synthesis of both short and long oligonucleotides. The mutant Y639F retains full enzymatic activity although individual kinetic parameters are somewhat different. Kinetic parameters, (kcat)app and (Km)app for the nucleotides, reveal that the mutation of Lys to Met has a much more drastic effect on (kcat)app than on (Km)app, indicating the involvement of K631 primarily in phosphodiester bond formation. The mutation of His to Gln has effects on both (kcat)app and (Km)app; namely, three- to fivefold reduction in (kcat)app and two- to threefold increase in (Km)app, implying that His811 may be involved in both nucleotide binding and phosphodiester bond formation. The ability of the mutant T7 RNA polymerases to bind template has not been greatly impaired. We have shown that amino acids D537 and D812 are essential, that amino acids K631 and H811 play significant roles in catalysis, and that the active site of T7 RNA polymerase is composed of different regions of the polypeptide chain. Possible roles for these catalytically significant residues in the polymerase mechanism are discussed. 相似文献
25.
Construction of a human cytochrome c gene and its functional expression in Saccharomyces cerevisiae 总被引:1,自引:0,他引:1
Y Tanaka T Ashikari Y Shibano T Amachi H Yoshizumi H Matsubara 《Journal of biochemistry》1988,103(6):954-961
The nucleotide sequences of a partial cDNA and three pseudogenes of human cytochrome c were determined. The complete nucleotide sequences which encode human cytochrome c were constructed on the basis of one of the pseudogenes by in vitro mutagenesis. The constructed human cytochrome c was functionally expressed in Saccharomyces cerevisiae. The recombinant human cytochrome c was purified and characterized. 相似文献
26.
27.
L-myo-Inositol-1-phosphate synthase has been found to have at least a 5-fold preference for the beta-anomer of its natural substrate D-Glc-6-P. The alpha-anomer appears to be an inhibitor of the reaction and may be converted to product as well. As well as showing an enzymatic preference for the equatorial C-1 hydroxyl of D-Glc-6-P, our results suggest that it is the pyranose form of D-Glc-6-P that binds to the enzyme and that ring-opening is an enzymatic step. We have also found D-2-dGlc-6-P, D-2-F-2-dGlc-6-P, and D-Man-6-P each to be both competitive inhibitors and substrates that are converted to inositol phosphates by the synthase. D-Allose-6-P is a weak inhibitor of the enzyme, but not a substrate. D-Gal-6-P is neither substrate nor inhibitor. Thus the specificity of the synthase with respect to single position epimers of D-Glc-6-P increases in the order C1 less than C2 much less than C3 less than C4. 相似文献
28.
F. Kato T. Hino A. Nakaji M. Tanaka Y. Koyama 《Molecular & general genetics : MGG》1995,247(3):387-390
In many species of actinomycetes, carotenogenesis can be photoinduced. The capacity to respond to photoinduction is, however unstable and, in various strains of Streptomyces, is lost at a relatively high frequency. In Streptomyces setonii ISP5395, which normally produces no carotenoids, carotenoid-producing mutants can be obtained following protoplast regeneration. We report here the characterization of a gene, crtS, which was isolated from one such mutant and can confer on wild-type S. setonii ISP5395 cells the capacity to synthesize carotenoids. Sequence analysis of crtS reveals an open reading frame, which shows homology to genes that encode alternative sigma factors in Bacillus subtilis. We propose that crtS encodes a sigma factor which is necessary for the expression of a cryptic gene(s) for carotenoid biosynthesis in S. setonii ISP5395. 相似文献
29.
30.