全文获取类型
收费全文 | 3116篇 |
免费 | 202篇 |
国内免费 | 1篇 |
出版年
2022年 | 19篇 |
2021年 | 35篇 |
2020年 | 25篇 |
2019年 | 35篇 |
2018年 | 33篇 |
2017年 | 21篇 |
2016年 | 58篇 |
2015年 | 68篇 |
2014年 | 81篇 |
2013年 | 331篇 |
2012年 | 182篇 |
2011年 | 153篇 |
2010年 | 98篇 |
2009年 | 116篇 |
2008年 | 139篇 |
2007年 | 141篇 |
2006年 | 134篇 |
2005年 | 108篇 |
2004年 | 131篇 |
2003年 | 127篇 |
2002年 | 106篇 |
2001年 | 87篇 |
2000年 | 114篇 |
1999年 | 111篇 |
1998年 | 34篇 |
1997年 | 25篇 |
1996年 | 23篇 |
1995年 | 21篇 |
1994年 | 16篇 |
1993年 | 23篇 |
1992年 | 66篇 |
1991年 | 61篇 |
1990年 | 52篇 |
1989年 | 38篇 |
1988年 | 55篇 |
1987年 | 32篇 |
1986年 | 41篇 |
1985年 | 30篇 |
1984年 | 39篇 |
1983年 | 29篇 |
1982年 | 29篇 |
1981年 | 29篇 |
1980年 | 17篇 |
1979年 | 18篇 |
1978年 | 19篇 |
1977年 | 19篇 |
1974年 | 18篇 |
1973年 | 16篇 |
1972年 | 19篇 |
1967年 | 18篇 |
排序方式: 共有3319条查询结果,搜索用时 359 毫秒
31.
Early replicative intermediates of Escherichia coli chromosome isolated from a membrane complex. 总被引:5,自引:3,他引:2 下载免费PDF全文
The replication origin region of the Escherichia coli chromosome was isolated from an outer membrane fraction. The chromosome was further purified by centrifugation in a cesium chloride gradient. Early replicative intermediates were enriched in the preparation when cytosine-1-beta-arabinofuranoside was added to the culture at the time of initiation of chromosome replication. DNA fragments with an eye structure having two branches of less than 400-500 bp in length were associated with components that were removed by phenol treatment. We conclude that the replication fork usually proceeds counter-clockwise toward the unc operon in the earliest period of replication. 相似文献
32.
Investigation of actin in Tetrahymena cells. A comparison with skeletal muscle actin by a devised two-dimensional gel electrophoresis method 总被引:2,自引:0,他引:2
Total protein constituents of Tetrahymena thermophila strain B1868 III were studied by two-dimensional agarose-polyacrylamide gel electrophoresis to detect actin among the constituents. In the attempts to prepare a whole-cell extract of Tetrahymena, it was found that protease activity in the extract was so high that high molecular components were quickly digested with the endogenous protease into small peptides unless the homogenization and heat-treatment in a sodium dodecylsulfate solution were performed within 5 s. It was eventually found that employment of 8 M guanidine hydrochloride (HCl) in the homogenization of cells perfectly prevented the degradation of protein components, even through a long preparation procedure. A devised two-dimensional agarose-polyacrylamide gel electrophoresis of the guanidine HCl extract gave a 'protein map' on which most proteins were located in their respective positions, including proteins with more than 200,000 mol. wt. Addition of rabbit skeletal muscle actin on the protein map revealed that no protein with isoelectric point and molecular weight identical with those of the actin was contained in the whole Tetrahymena extract, suggesting that Tetrahymena actin may have characteristics far different from those of skeletal muscle actin. 相似文献
33.
Spirulina ferredoxin-NADP+ reductase. Further characterization with an improved preparation 总被引:2,自引:0,他引:2
The preparation procedure for Spirulina ferredoxin-NADP+ reductase (ferredoxin: NADP+ oxidoreductase, EC 1.18.1.2, FNR) was improved by adding protease inhibitors, phenylmethylsulfonylfluoride (PMSF) and EDTA, through the whole process of preparation and by introducing an affinity chromatography step on Blue Sepharose CL-6B. The addition of the inhibitors largely prevented the formation of the minor component (FNR I), and the affinity gel chromatography simplified the preparation process, shortening the exposure period of FNR to proteolysis. However, complete removal of the heterogeneity of FNR found at the amino (N)-terminal region was not achieved even by applying the new method. The affinity chromatography on the Blue Sepharose gel was also effective in purifying spinach FNR. The affinity of this gel for Spirulina FNR was compared with that for the enzyme derived from spinach leaves. The spinach enzyme had a higher affinity than the Spirulina one. Both enzymes showed the highest affinities to Blue Sepharose at 20--30 mM NaCl concentration. The N-terminal sequence analysis revealed that there was 4 forms, which were probably modifications produced by exopeptidase action during the preparation, or even in the living cells. The longest component gave the N-terminal sequence Ala-Lys-Thr-Asp-Ile-Pro-Val-Asn-Ile-Tyr-. The others lacked amino acids successively one by one from the N-terminus. In contrast, the carboxyl(C)-terminal residues of all 4 FNR forms were tyrosine. The probable C-terminal sequence was predicted to be -Trp-His-Val-Gln-Thr-Tyr based on a study of a cyanogen bromide peptide. 相似文献
34.
Pleural effusions and sera of two patients with lung cancer were tested after intrapleural injection of OK-432 as an anticancer drug for IFN-alpha activity by biological assay and for IFN-alpha as an antigen by radioimmunoassay. The titers by radioimmunoassay were fairly consistent with those by biological assay, but were usually higher. In Case 1, IFN-alpha was observed fairly early after administration of OK-432 and only in pleural effusions. In Case 2, induction of IFN-alpha at low level was observed late after the first administration of OK-432 both in the pleural effusion and serum and was detected only by radioimmunoassay. 相似文献
35.
A role of the B-oligomer moiety of islet-activating protein, pertussis toxin, in development of the biological effects on intact cells 总被引:52,自引:0,他引:52
Islet-activating protein (IAP), pertussis toxin, is an oligomeric protein (Tamura, M., Nogimori, K., Murai, S., Yajima, M., Ito, K., Katada, T., Ui, M., and Ishii, S. (1982) Biochemistry 21, 5516-5522), the biggest subunit (Mr = 28,000, referred to as the A-protomer) of which catalyzes transfer of the ADP-ribose moiety of NAD to the membrane Mr = 41,000 protein. The pentamer, termed the B-oligomer, consisting of the residual subunits was the moiety of IAP that was responsible for binding to the cell surface, as revealed by competitive inhibition of the development of the IAP actions on intact rat C6 glioma cells and rat adipocytes. The binding of the B-oligomer to its receptor proteins was divalent via the constituent two dimers; it stimulated mitosis of lymphocytes and caused an insulin-like action to enhance glucose oxidation in adipocytes, just as did concanavalin A, presumably as a result of cross-linking or aggregation of the membrane proteins. The A-promoter displayed its biological action on adipocytes only when the B-oligomer had been bound to the cells. Thus, IAP is a typical A-B toxin in which the B-oligomer is first bound to the cell surface proteins to enable the A-protomer to reach to the site of its action within the cell. Diverse biological actions of pertussis toxin may be accounted for by the mitogenic action of the B-oligomer as well as ADP-ribosyltransferase activity of the A-promoter. 相似文献
36.
Effects of salt and pH on the re-reduction of P700 by chemically-modifiedhorse heart cytochrome c after a flash illumination were examinedin Triton-treated P700- enriched subchloroplast particles (TSF-1particles). At low salt concentrations net charges on the membrane surfaceand native, guanidinated or succinylated cytochrome c were majorfactors that determined the reaction rates, as in the reactionbetween plastocyanin and P700 [Tamura et al. (1981) Plant &Cell Physiol. 22: 603]. The reaction rates also depended onreactant-specific factors, particularly the localized distributionof charges on macromolecules and their interaction over shortdistances, as well as on long-range Coulombic interaction. Theeffect of this type became clearer at high salt concentrations. (Received October 7, 1982; Accepted December 20, 1982) 相似文献
37.
Separation of the Polypeptides of Chlamydia and Its Cell Walls by Polyacrylamide Gel Electrophoresis 总被引:6,自引:3,他引:3 下载免费PDF全文
The polypeptide composition of Chlamydia was examined by acrylamide gel electrophoresis. When the polypeptide patterns of purified infectious elementary bodies (EB) of C. psittaci meningopneumonitis strain, 6BC strain, and C. trachomatis T'ang strain were compared, no significant differences were observed. The polypeptide patterns of whole EB and reticulate bodies (RB) appeared to overlap, but differences were found. In EB cell walls, nine main and several minor bands of polypeptides were observed in gels containing sodium lauryl sulfate, and the eighth main band from the top of the gel stained positive with periodic acid-Schiff reagent. On the other hand, the polypeptides in bands 3, 6, and 8 in EB cell walls were missing or minor in RB cell walls, and the ninth band was clearly stained by PAS. Band 8 was also stained slightly. Purified subunits, which occur as a lattice structure on the inside layer of EB cell walls but are largely missing in RB cell walls, contained bands 4, 6, and 8, and band 8 was PAS positive. These results indicate that significant polypeptide synthesis or reorganization in the cell walls occurs during the growth cycle. 相似文献
38.
39.
40.
A Tamura 《Journal of bacteriology》1967,93(6):2009-2016
In ribonucleic acid (RNA) extracted by phenol and sodium dodecyl sulfate from purified reticulate bodies of meningopneumonitis (MP) organisms, 21S, 16S, and 4S RNA were found by sucrose density gradient sedimentation analysis. When purified reticulate bodies were homogenized by sonic treatment or by treatment with sodium deoxycholate and were fractionated by differential centrifugation, more than 50% of the RNA was recovered in the fraction which was sedimented by centrifugation at 105,000 x g for 2 hr, but not at 13,000 x g for 20 min. From homogenates prepared in this manner, 50S and 30S particles containing RNA were isolated by sucrose density gradient centrifugation. These 50S and 30S particles were also found in lysates of cytoplasmic fractions of infected cells which were labeled by (32)P during 17 to 17.5 hr or 15 to 18 hr after infection. The synthesis of 50S and 30S particles was not inhibited by actinomycin D. When infected cells were homogenized in the presence of 0.01 or 0.02 m MgCl(2), 70S particles were isolated instead of 50S and 30S particles. When dialyzed against low concentrations of MgCl(2), the 70S particles dissociated to 50S and 30S particles. The base ratio of the 70S particles is very similar to that of 16S plus 21S RNA. The characteristics of the 70S, 50S, and 30S particles suggest that these are ribosome particles, similar to bacterial ribosomes. 相似文献