全文获取类型
收费全文 | 382篇 |
免费 | 35篇 |
国内免费 | 1篇 |
专业分类
418篇 |
出版年
2022年 | 7篇 |
2021年 | 7篇 |
2020年 | 5篇 |
2019年 | 8篇 |
2018年 | 3篇 |
2017年 | 6篇 |
2016年 | 4篇 |
2015年 | 14篇 |
2014年 | 14篇 |
2013年 | 14篇 |
2012年 | 21篇 |
2011年 | 14篇 |
2010年 | 15篇 |
2009年 | 12篇 |
2008年 | 20篇 |
2007年 | 18篇 |
2006年 | 22篇 |
2005年 | 18篇 |
2004年 | 5篇 |
2003年 | 23篇 |
2002年 | 12篇 |
2001年 | 7篇 |
2000年 | 5篇 |
1999年 | 8篇 |
1998年 | 2篇 |
1995年 | 5篇 |
1994年 | 2篇 |
1993年 | 2篇 |
1992年 | 5篇 |
1991年 | 7篇 |
1990年 | 9篇 |
1989年 | 13篇 |
1988年 | 12篇 |
1987年 | 11篇 |
1986年 | 5篇 |
1985年 | 5篇 |
1984年 | 6篇 |
1983年 | 4篇 |
1982年 | 5篇 |
1981年 | 2篇 |
1975年 | 1篇 |
1972年 | 2篇 |
1969年 | 1篇 |
1968年 | 1篇 |
1965年 | 1篇 |
1964年 | 1篇 |
1954年 | 2篇 |
1952年 | 10篇 |
1951年 | 11篇 |
1950年 | 8篇 |
排序方式: 共有418条查询结果,搜索用时 0 毫秒
1.
In this paper the recent population changes of the Wild Boar in different European countries is analysed through the study of hunting statistics. A simultaneous increase in numbers is observed throughout the whole area during the period 1965–1975. From 1975 onwards the population stabilizes itself apart from in peripheral areas like Finland. Potentially favourable factors which play a part in this process are discussed and certain reproductive and dispersive characteristics which favour its invasive behaviour are discussed. 相似文献
2.
Affinity purification of the HIV-1 protease 总被引:5,自引:0,他引:5
J C Heimbach V M Garsky S R Michelson R A Dixon I S Sigal P L Darke 《Biochemical and biophysical research communications》1989,164(3):955-960
An inhibitor of the HIV-1 protease has been employed in the generation of a resin which allows the rapid purification of this enzyme. A peptide substrate analogue, H2N-Ser-Gln-Asn-(Phe-psi[CH2N]-Pro)-Ile-Val-Gln-OH, was coupled to agarose resin. The HIV-1 protease was expressed in E. coli and the supernatant from lysed cells was passed through the affinity resin. Active HIV-1 protease was then eluted with a buffer change to pH 10 and 2 M NaCl. Final purification to a homogeneous preparation, capable of crystallization, was achieved with hydrophobic interaction chromatography. Solutions containing HIV-1 protease bound to competitive inhibitors do not bind to the column. 相似文献
3.
Monoclonal antibody Y13-259 to ras p21 was shown to bind to the highly conserved residues in the region 63-73 and to neutralize ras action in the Saccharomyces cerevisiae adenylate cyclase system. Inhibition of adenylate cyclase activity in isolated membranes by antibody Y13-259 occurred after a lag period of 6 min. This lag corresponded to the time necessary for binding of antibody Y13-259 to the membranes in a ras-dependent manner. The mechanism of inhibition appeared to be steric in nature because antibody Y13-259 neutralized ras p21 bound to a stable GTP analogue. Monoclonal antibodies Y13-4 and Y13-128 also inhibited yeast adenylate cyclase activity, and the epitopes for both the these antibodies were localized to ras region 65-75. However, the ras residues essential for binding of antibodies Y13-4 and Y13-128 to ras p21 (positions 65, 66, 68 and 75) were different from those essential for binding of antibody Y13-259 (positions 63, 65, 66, 67, 70 and 73). These results indicate that residues 63-75 constitute a major neutralizing epitope on ras p21. 相似文献
4.
5.
G E Rukavishnikova M I Potapov E R Sigal 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1985,(1):69-73
The use of the indirect ELISA techniques did not ensure the sharp differentiation of the antigens of the blood groups A and B on the polystyrene sorbent by means of heteroimmune sera, though such differentiation could be achieved by means of monoclonal antibodies. The test system known as "the lectin-antibody sandwich" was found to have the optimum sensitivity and specificity permitting the detection of soluble ABH antigens. This variant of ELISA permitted the detection of blood group A antigen both in native biological materials and in traces of blood and saliva, thus making it possible to carry out its quantitative determination. 相似文献
6.
Patterns of isotype commitment in human B cells: limiting dilution analysis of Epstein Barr virus-infected cells 总被引:4,自引:0,他引:4
L D Stein C J Ledgley N H Sigal 《Journal of immunology (Baltimore, Md. : 1950)》1983,130(4):1640-1645
The frequency of Epstein Barr virus- (EBV) inducible IgM, IgG, and IgA-secreting cells in human peripheral blood and tonsil was determined by performing limiting dilution experiments in suspension culture. We devised a method of propagating small numbers of EBV-infected B cells with irradiated umbilical cord blood cells as a feeder layer. Precursor cell frequencies can be derived from these experiments; we have shown by statistical analysis that they conform to the single hit model of the Poisson distribution. By using this technique, a significant percentage of surface immunoglobulin-bearing lymphocytes are activated to secrete immunoglobulin in vitro. On the average, 8 to 38% of B cells in peripheral blood and tonsil can be propagated and the secreted immunoglobulin from the clonal progeny can be analyzed. Neither the EBV immune status of the donor nor the source of the umbilical cord blood feeder layer could account for the variations in cloning efficiency observed among donors. A surprisingly high frequency of B cells secreted IgA in vitro and we have shown that a small proportion of B cell clones in tonsil and peripheral blood secrete both IgM and IgA during the 4-wk culture period. Other B cells, including all those that produce IgG, appear to be committed to the secretion of a single isotype. Thus, these studies establish methodology for the analysis of the secreted products of human B cells at the single cell level and demonstrate that the progeny of at least some clones can secrete more than one isotype in vitro. 相似文献
7.
Ras interaction with the GTPase-activating protein (GAP) 总被引:18,自引:0,他引:18
M D Schaber V M Garsky D Boylan W S Hill E M Scolnick M S Marshall I S Sigal J B Gibbs 《Proteins》1989,6(3):306-315
Biologically active forms of Ras complexed to GTP can bind to the GTPase-activating protein (GAP), which has been implicated as possible target of Ras in mammalian cells. In order to study the structural features of Ras required for this interaction, we have evaluated a series of mutant ras proteins for the ability to bind GAP and a series of Ras peptides for the ability to interfere with this interaction. Point mutations in the putative effector region of Ras (residues 32-40) that inhibit biological activity also impair Ras binding to GAP. An apparent exception is the Thr to Ser substitution at residue 35; [Ser-35]Ras binds to GAP as effectively as wild-type Ras even though this mutant is biologically weak in both mammalian and S. cerevisiae cells. In vitro, [Ser-35]Ras can also efficiently stimulate the S. cerevisiae target of Ras, adenylyl cyclase, indicating that other factors may influence Ras/protein interactions in vivo. Peptides having Ras residues 17-44 and 17-32 competed with the binding of Ras to E. coli-expressed GAP with IC50 values of 2.4 and 0.9 microM, respectively, whereas Ras peptide 17-26 was without effect up to 400 microM. A related peptide from the yeast GTP-binding protein YPT1 analogous to Ras peptide 17-32 competed with an IC50 value of 19 microM even though the YPT1 protein itself is unable to bind to GAP. These results suggest that determinants within Ras peptide 17-32 may be important for Ras binding to GAP. 相似文献
8.
A site-specific mutation within the active site of ribulose-1,5-bisphosphate carboxylase of Rhodospirillum rubrum 总被引:8,自引:0,他引:8 下载免费PDF全文
In vitro mutagenic techniques have generated an asp→glu substitution at residue 198 adjacent to the carbamate-divalent metal ion binding site of Rhodospirillum rubrum ribulose 1,5-bisphosphate carboxylase. A single C→A nucleotide change in the coding strand created the mutant and introduced a new EcoRI restriction site on the expression plasmid pRR2119. Although the carboxylase:oxygenase ratio remained the same, the mutant enzyme had slightly altered kinetic properties. The e.p.r. spectra of the quaternary complexes enzyme.activator carbamate.Mn2+.2-carboxyarabinitol 1,5-bisphosphate and enzyme.activator carbamate.Mn2+.4-carboxyarabinitol 1,5-bisphosphate for mutant and wild-type enzymes were different, indicating that the metal ion was in a slightly altered environment. These findings are consistent with the hypothesis that, besides the carbamate at lys 201, the carboxyl group of asp 198 contributes to the formation of the divalent metal ion binding site. 相似文献
9.
FK-506 and cyclosporin A inhibit highly similar signal transduction pathways in human T lymphocytes. 总被引:7,自引:0,他引:7
This report compares the ability of cyclosporin A and FK-506 to inhibit human T cell activation triggered via cell surface molecules that utilize different intracellular processes. We stimulated highly purified peripheral blood T lymphocytes with mitogens (Con A and PHA), ionomycin + PMA, or monoclonal antibodies specific for cell surface antigens involved in activation (CD2, CD3, CD28) either in combination with each other or in conjunction with PMA. Using measurements of the proliferative response, IL-2 production, and changes in intracellular Ca2+ ([Ca2+]i), we demonstrate that FK-506 exerts its inhibitory effect on early events of T-cell activation in a manner indistinguishable from that of CsA. An important finding in this study is the strict correlation between those activation pathways that are inhibited by FK-506 and CsA and the requirement that the sensitive pathways induce a measurable rise in [Ca2+]i. This correlation held even for the CD28/CD2 pathway which was previously shown to be calcium-independent; however by employing FACS analysis of [Ca2+]i within individual cells, a subset of cells activated via CD28/CD2 was found to respond with a measurable rise in [Ca2+]i. We also noted that the proliferative response induced by certain stimuli, such as ionomycin + PMA and PHA + PMA, was partially resistant to FK-506 and CsA, while IL-2 production was completely suppressed. The partial FK-506/CsA-resistance of these responses was shown to be determined by the amount of PMA added to the cultures. We conclude from our investigations that FK-506 and CsA inhibit highly similar signal transduction pathways in human T lymphocytes. 相似文献
10.
Tslil Gabrieli Yael Michaeli Sigal Avraham Dmitry Torchinsky Sapir Margalit Leonie Schütz Matyas Juhasz Ceyda Coruh Nissim Arbib Zhaohui Sunny Zhou Julie
A Law Elmar Weinhold Yuval Ebenstein 《Nucleic acids research》2022,50(16):e92
DNA methylation, specifically, methylation of cytosine (C) nucleotides at the 5-carbon position (5-mC), is the most studied and significant epigenetic modification. Here we developed a chemoenzymatic procedure to fluorescently label non-methylated cytosines in CpG context, allowing epigenetic profiling of single DNA molecules spanning hundreds of thousands of base pairs. We used a CpG methyltransferase with a synthetic S-adenosyl-l-methionine cofactor analog to transfer an azide to cytosines instead of the natural methyl group. A fluorophore was then clicked onto the DNA, reporting on the amount and position of non-methylated CpGs. We found that labeling efficiency was increased up to 2-fold by the addition of a nucleosidase, presumably by degrading the inactive by-product of the cofactor after labeling, preventing its inhibitory effect. We used the method to determine the decline in global DNA methylation in a chronic lymphocytic leukemia patient and then performed whole-genome methylation mapping of the model plant Arabidopsis thaliana. Our genome maps show high concordance with published bisulfite sequencing methylation maps. Although mapping resolution is limited by optical detection to 500–1000 bp, the labeled DNA molecules produced by this approach are hundreds of thousands of base pairs long, allowing access to long repetitive and structurally variable genomic regions. 相似文献