全文获取类型
收费全文 | 512篇 |
免费 | 63篇 |
专业分类
575篇 |
出版年
2023年 | 2篇 |
2022年 | 6篇 |
2021年 | 7篇 |
2019年 | 9篇 |
2018年 | 9篇 |
2017年 | 17篇 |
2016年 | 18篇 |
2015年 | 14篇 |
2014年 | 12篇 |
2013年 | 41篇 |
2012年 | 39篇 |
2011年 | 42篇 |
2010年 | 15篇 |
2009年 | 15篇 |
2008年 | 33篇 |
2007年 | 21篇 |
2006年 | 19篇 |
2005年 | 21篇 |
2004年 | 17篇 |
2003年 | 18篇 |
2002年 | 9篇 |
2001年 | 17篇 |
2000年 | 22篇 |
1999年 | 18篇 |
1998年 | 2篇 |
1997年 | 7篇 |
1996年 | 4篇 |
1995年 | 5篇 |
1994年 | 7篇 |
1993年 | 2篇 |
1992年 | 13篇 |
1991年 | 16篇 |
1990年 | 10篇 |
1989年 | 13篇 |
1988年 | 5篇 |
1987年 | 7篇 |
1986年 | 7篇 |
1985年 | 5篇 |
1984年 | 5篇 |
1983年 | 6篇 |
1981年 | 3篇 |
1979年 | 3篇 |
1978年 | 2篇 |
1973年 | 1篇 |
1972年 | 1篇 |
1970年 | 3篇 |
1968年 | 2篇 |
1967年 | 1篇 |
1966年 | 1篇 |
1962年 | 1篇 |
排序方式: 共有575条查询结果,搜索用时 15 毫秒
1.
S Papa M Vitale G Mazzotti R Rizzoli M Falconi A Bartoletti F A Manzoli 《Histochemistry》1988,89(3):241-245
A number of techniques are currently employed for the fractionation of heterogeneous cell populations or for the separation of cells in different phases of their cycle. With the development of osmotically inert colloidal silica particles media, density gradient centrifugation became an established method for the separation and purification of cells and subcellular particles. We have applied this technique to the separation of cycling from resting Friend erythroleukemia cells, to obtain purified populations for further biological assays. The flow cytometric analysis of DNA content of the different fractions obtained by the gradient and stained with Propidium Iodide (PI), showed the S compartment highly concentrated in the 1.073/77 g/ml interface, while the upper levels of the gradient were highly enriched of cells in G1 phase. Moreover, the dual parameter analysis of DNA content by means of Bromodeoxyuridine (BrdUrd) incorporation and PI staining, showed that part of the cells in the 1.067/73 fraction represented the early S phase even if their DNA level, measured on the basis of PI fluorescence was within the diploid cell cluster. This method seems to be suitable to obtain pure cell fractions even when dealing with numerically large populations. 相似文献
2.
S Papa S Capitani A Matteucci M Vitale P Santi A M Martelli N M Maraldi F A Manzoli 《Cytometry》1987,8(6):595-601
The development of hepatocyte polyploidy in rats aged up to 4 months was analyzed by flow cytometry using both scatter and fluorescent parameters to distinguish DNA diploid and DNA tetraploid populations and to discriminate between parenchymal and non-parenchymal compartments. The precise origin of each class of nuclei was assessed in whole liver homogenate using purified hepatocytes, obtained by liver perfusion followed by separation on Percoll gradient, and identifying the peaks corresponding to parenchymal nuclei. The results indicate that preparative procedures involving homogenization of the rat liver tissue caused loss of the DNA octaploid population. Data on the relative proportion of the different DNA ploidy elements during rat liver development, which are in good agreement with those observed by cell analysis by means of microspectrophotometry, indicate the usefulness of flow cytometry as a choice method for the analysis of ploidy distribution. 相似文献
3.
Summary A trypsin-like proteinase was isolated from Streptomyces rimosus culture filtrates obtained from an oxytetracycline production process. The isolation procedure includes ultrafiltration, chromatography on CM-Sephadex, AH-Sepharose and CM-cellulose and gives a homogeneous protein with 19% yield. The enzyme is an anionic trypsin (Mr 28 000, pI 4.5), is stable from pH 4.5 to 9 and up to 40°C, and contains three disulphide bridges, three histidines and three methionines per molecule. At its pH optimum (pH 8.4–8.8) it splits peptide, ester and arylamide bonds of arginine in the endo-position and, to a smaller extent, in the exo-position. Like other streptomycete trypsins, it is a more efficient catalyst than bovine trypsin and has a relative preference for peptide-arylamides, N-benzyloxycarbonyl-l-norleucyl-l-prolyl-l-arginine-p-nitroanilide being by far its best substrate. 相似文献
4.
S Petrovi? L Vitale 《Comparative biochemistry and physiology. B, Comparative biochemistry》1990,95(3):589-595
Hydrolytic activities characteristic for different aminopeptidases were detected in the egg-white of unfertilized chicken eggs, and one aminopeptidase was isolated in an electrophoretically homogeneous form. The isolated aminopeptidase preferentially hydrolyzed bonds of alpha-glutamyl residue at the NH(2)-end of synthetic substrates and peptides. The enzyme is a dimer with an M(r) of 320,000 and pI of 4.2. Its optimal pH and temperature are 7.6 and 60 degrees C, respectively. EDTA, amastatin, and N-bromosuccinimide are inhibitors, while Ca2++ and Mn2+ are activators of the enzyme Ca2+ also stabilizes the enzyme. According to the observed properties, the isolated chicken egg-white aminopeptidase belongs to the glutamyl aminopeptidases. 相似文献
5.
Lj. Vitale M. Renko B. Lenarčič V. Turk M. Pokorny 《Applied microbiology and biotechnology》1986,23(6):449-455
Summary A leucine aminopeptidase was purified to homogeneity fromStreptomyces rimosus culture filtrates, which are waste broth of oxytetracycline bioproduction process. Purification procedure includes ultrafiltration
and chromatography on CM-Sephadex, AH-Sepharose and FPLC Mono S column.
The enzyme is a monomer with molecular weight of 27,500 Daltons and pI of 7.3, stable in broad pH range and up to 70°C. It
is a metallo enzyme dependent on Ca2+ ions for its full activity. By its specificity it is a true aminopeptidase active on amino acid amide, arylamide, peptide
and ester bonds. The hydrolysing activity shows preference for leucine at the N-terminal position of substrates, also acts
on aromatic acids and methionine, but does not release glycine, proline, acidic amino acids orD-amino acid residues. 相似文献
6.
7.
Binding of BiP to an assembly-defective protein in plant cells 总被引:5,自引:1,他引:4
Emanuela Pedrazzini Giovanna Giovinazzo Roberto Bollini Aldo Ceriotti Alessandro Vitale 《The Plant journal : for cell and molecular biology》1994,5(1):103-110
The binding protein (BiP) has been implicated as a mediator of protein folding and assembly in the endoplasmic reticulum of mammalian cells and has often been found in stable association with structurally defective proteins. To acquire information on the activity of BiP in plant cells, we have expressed in tobacco protoplasts the wild type form and an assembly-defective form of bean phaseolin. Phaseolin (PHSL) is a soluble, trimeric, storage glycoprotein co-translationally inserted into the lumen of the endoplasmic reticulum and then transported along the secretory pathway to the protein storage vacuoles. We have previously shown that a PHSL mutant in which the last 59 amino acids have been deleted (Δ363PHSL) is unable to form trimers and is retained in a pre-Golgi compartment when synthesized in Xenopus oocytes. When transiently expressed in tobacco leaf protoplasts, wild-type PHSL is correctly glycosylated and assembles efficiently and rapidly into trimers. Δ363PHSL is also correctly glycosylated but does not trimerize. Tobacco BiP and Δ363PHSL are co-immunoselected using either anti-PHSL or anti-BiP antibodies. Under the same conditions, co-immunoselection of BiP with wild-type PHSL is not detectable. The BiP bound to Δ363PHSL can be released by treatment of the complex with ATP, indicating that the binding is related to the proposed function of BiP in protein folding and assembly in the endoplasmic reticulum. These data indicate that BiP stably binds structurally defective proteins in plant cells. 相似文献
8.
From filtrates of an oxytetracycline-producing culture of Streptomyces rimosus a deoxyribonuclease was purified to homogeneity and determined to be a potent endo-DNase. It is a monomeric, basic protein (Mr 21 000; pI 9.5) stable in a broad pH range but unstable to higher temperature. The enzyme has an absolute requirement for Mg2 + or Mn2 +, and for its full activity requires free SH groups and a low-ionic-strength environment. Its N-terminal primary structure differs from that of other nucleases. 相似文献
9.
Blattella germanica (L.) that were fed hydramethylnon bait produced residues that were toxic to exposed conspecifics. Insecticidal activity was traced to the feces of treated insects by feeding radiolabeled material, where approximately 50% of the recovered radioactivity was unmetabolized parent compound. Ingestion of toxicant-laden feces by all life stages was evident, but the effect of this behavior was greatest on early instar nymphs. Baits containing toxicants with delayed activity, such as hydramethylnon, probably affect cockroach field populations indirectly through coprophagy. 相似文献
10.
Dr. Darrell R. Stokes Anthony J. Vitale Charles R. Morgan 《Cell and tissue research》1979,198(1):175-189
Histochemical techniques have been employed to characterize enzymatic activity in the mesocoxal muscles of the cockroach, Periplaneta americana. Through our studies of the enzymes myosin-ATPase, NADH reductase, succinic dehydrogenase (SDH), and lactic dehydrogenase (LDH), we were able to classify fibers within these muscles according to criteria established for muscle fibers of vertebrates. Many of the mesocoxal muscles possess two different and distinct populations of fibers, whereas the remaining muscles are homogeneous with respect to their constituent fibers. The data presented here indicate biochemical heterogeneity for muscles of differing structural and functional features and possible neurotrophic influences upon oxidative enzymes and myosin-ATPase isozymes. 相似文献