全文获取类型
收费全文 | 8666篇 |
免费 | 727篇 |
国内免费 | 1篇 |
出版年
2023年 | 51篇 |
2022年 | 109篇 |
2021年 | 234篇 |
2020年 | 151篇 |
2019年 | 167篇 |
2018年 | 193篇 |
2017年 | 181篇 |
2016年 | 293篇 |
2015年 | 464篇 |
2014年 | 503篇 |
2013年 | 602篇 |
2012年 | 756篇 |
2011年 | 712篇 |
2010年 | 435篇 |
2009年 | 407篇 |
2008年 | 560篇 |
2007年 | 496篇 |
2006年 | 462篇 |
2005年 | 406篇 |
2004年 | 355篇 |
2003年 | 305篇 |
2002年 | 319篇 |
2001年 | 89篇 |
2000年 | 84篇 |
1999年 | 101篇 |
1998年 | 52篇 |
1997年 | 49篇 |
1996年 | 47篇 |
1995年 | 42篇 |
1994年 | 39篇 |
1993年 | 31篇 |
1992年 | 54篇 |
1991年 | 50篇 |
1990年 | 50篇 |
1989年 | 51篇 |
1988年 | 30篇 |
1987年 | 41篇 |
1986年 | 41篇 |
1985年 | 38篇 |
1984年 | 30篇 |
1983年 | 25篇 |
1982年 | 23篇 |
1981年 | 23篇 |
1980年 | 20篇 |
1979年 | 17篇 |
1978年 | 20篇 |
1977年 | 24篇 |
1976年 | 20篇 |
1975年 | 21篇 |
1973年 | 28篇 |
排序方式: 共有9394条查询结果,搜索用时 968 毫秒
71.
Cloning and characterization of human immunodeficiency virus type 1 variants diminished in the ability to induce syncytium-independent cytolysis. 总被引:7,自引:5,他引:2
下载免费PDF全文
![点击此处可从《Journal of virology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
M Stevenson S Haggerty C Lamonica A M Mann C Meier A Wasiak 《Journal of virology》1990,64(8):3792-3803
The phenomenon of interference was exploited to isolate low-abundance noncytopathic human immunodeficiency virus type 1 (HIV-1) variants from a primary HIV-1 isolate from an asymptomatic HIV-1-seropositive hemophiliac. Successive rounds of virus infection of a cytolysis-susceptible CD4+ cell line and isolation of surviving cells resulted in selective amplification of an HIV-1 variant reduced in the ability to induce cytolysis. The presence of a PvuII polymorphism facilitated subsequent amplification and cloning of cytopathic and noncytopathic HIV-1 variants from the primary isolate. Cloned virus stocks from cytopathic and noncytopathic variants exhibited similar replication kinetics, infectivity, and syncytium induction in susceptible host cells. The noncytopathic HIV-1 variant was unable, however, to induce single-cell killing in susceptible host cells. Construction of viral hybrids in which regions of cytopathic and noncytopathic variants were exchanged indicated that determinants for the noncytopathic phenotype map to the envelope glycoprotein. Sequence analysis of the envelope coding regions indicated the absence of two highly conserved N-linked glycosylation sites in the noncytopathic HIV-1 variant, which accompanied differences in processing of precursor gp160 envelope glycoprotein. These results demonstrate that determinants for syncytium-independent single-cell killing are located within the envelope glycoprotein and suggest that single-cell killing is profoundly influenced by alterations in envelope sequence which affect posttranslational processing of HIV-1 envelope glycoprotein within the infected cell. 相似文献
72.
Protein engineering by cDNA recombination in yeasts: shuffling of mammalian cytochrome P-450 functions 总被引:7,自引:0,他引:7
We have constructed, in the yeast Saccharomyces cerevisiae, a mosaic assembly of genes by in vivo recombination of partially homologous sequences. The approach was tested on cDNAs encoding functionally distinct mammalian cytochromes P-450 (P-450). The selection for recombinant cDNAs used the transformation of yeast cells, which required the recircularization of a linearized plasmid by recombination of two partially homologous cDNAs. Libraries of mosaic genes with bipartite or tripartite structures were generated by intramolecular and intermolecular recombination events. The presence of yeast promoter and terminator sequences on the flanking sides of the recombined cDNAs has allowed the synthesis of encoded mosaic proteins. A library of yeast clones producing recombinant mouse P-450 P1 and rabbit P-450 LM4 was screened using functional criteria to identify chimeras with shuffled substrate specificity. Restriction mapping of mosaic genes, biochemical analysis of the synthesized proteins, comparison of chimeric enzymes, and the alignment of sequences with bacterial P-450 camphor hydroxylase of known three-dimensional structure, all suggest that the P-450 P1 amino acid residues 203-238 play a major role in the control of cytochrome activity toward carcinogenic polycyclic aromatic hydrocarbons. Similar approaches to structure-function analysis are believed to be applicable to other protein families. 相似文献
73.
B. Meier 《Human Evolution》1989,4(2-3):223-229
Extinction of small, closed populations in captivity as well as in the wild is believed to be nearly inevitable, because inbreeding
will adversely effect reproductive success, mortality, sex ratio and also the susceptibility to epidemic diseases and environmental
stress. An ever increasing number of primate species exist only in small isolated populations, which contain only a part of
the original genetic variability. In captive breeding programs research about genetic management strategies is, therefore,
of essential importance. In 1980 we imported 9Loris tardigrdus nordicus (4 females, 5 males) from NE-Sri Lanka. The founders came from one natural breeding population. All sexual mature females
are breeding. Up to now the colony contains 36 living individuals. The main goal of our long-term genetic management plan
was to minimize inbreeding and to preserve the genetic diversity. Therefore, we try to pass the founder bottleneck rapidly
by enlarging the population to a desired minimum population size of 25 pairs and to equalize the founder representation within
any generation. The need to control the spread of sublethal genes, introduced by one of the founders, conflicts directly with
the aim of equalizing the founder representation. A solution of this problem is discussed. To produce a sufficiently large
population we intend to give animals to other institutions and to build up an exchange-system for offspring individuals, which
should lead to an international studbook. 相似文献
74.
M Repetto J C Maziere D Citadelle R Dupuis M Meier S Biade D Quiec C Roux 《Teratology》1990,42(6):611-618
AY 9944 [trans-1,4-bis(2-chlorobenzylaminomethyl) cyclohexane dihydrochloride] is an amphiphilic cationic molecule. This chemical is an established inhibitor of cholesterol synthesis and is teratogenic in rats. The mechanisms of this teratogenicity remain to be clarified. This study used cultured rat whole embryos to ascertain whether AY 9944 had a direct effect on embryos, or whether its action was indirect, via the maternal cholesterol metabolism. Four experimental conditions were investigated: (A) controls; (B) 10 day untreated embryos were cultured in serum of treated rats; (C) 10 day untreated embryos were cultured in serum containing added AY 9944 (0-1,000 micrograms/ml); and (D) 10 day embryos from females treated on day 4 of gestation were cultured in normal serum. In group B there was no growth retardation; some slight nonspecific abnormalities were not significant. In group C, direct addition of AY 9944 to culture medium retarded growth and differentiation in a dose-dependent manner. No malformation was observed, but histological examinations showed numerous areas of cell necrosis, especially in the CNS. In group D, not only was growth retardation observed, but also characteristic malformations of AY 9944 teratogenesis, including pituitary agenesis. These results show that AY 9944 teratogenicity is initiated prior to day 10. 相似文献
75.
Interaction of antimicrobial dermaseptin and its fluorescently labeled analogues with phospholipid membranes. 总被引:16,自引:0,他引:16
Dermaseptin, a 34 amino-acid residue antimicrobial polypeptide [Mor, A., Nguyen, V. H., Delfour, A., Migliore-Samour, D., & Nicolas, P. (1991) Biochemistry 30, 8824-8830] was synthesized and selectively labeled at its N-terminal amino acid with either 7-nitrobenz-2-oxa-1,3-diazole-4-yl (NBD), rhodamine, or fluorescein. The fluorescent emission spectra of the NBD-labeled dermaseptin displayed a blue-shift upon binding to small unilamellar vesicles (SUV), reflecting the relocation of the fluorescent probe to an environment of increased apolarity. Titrations of solutions containing NBD-labeled dermaseptin with SUV composed of zwitterionic or acidic phospholipids were used to generate binding isotherms, from which were derived surface partition constants of (0.66 +/- 0.06) x 10(4) M-1 and (2.8 +/- 0.3) x 10(4) M-1, respectively. The shape of the binding isotherms, as well as fluorescence energy transfer measurements, suggests that some aggregation of membrane-bound peptide monomers occurs in acidic but not in zwitterionic vesicles. The preferential susceptibility of the peptide to proteolysis when bound to zwitterionic but not to acidic SUV suggests that these aggregates might then penetrate a relatively short distance into the hydrophobic region of the acidic membrane. Furthermore, the results provide good correlation between the peptide's strong binding and its ability to permeate membranes composed of acidic phospholipids, as revealed by a dissipation of diffusion potential and a release of entrapped calcein from SUV. 相似文献
76.
E Boukhzer A Ennya F Felden A Gérard E Nexo J P Nicolas H Gérard J L Guéant 《Biochimica et biophysica acta》1992,1175(1):128-131
Specific binding sites for rabbit transcobalamin II have been found on isolated adult rabbit germ cells. Scatchard analysis revealed a single class of binding sites for [57Co]cyanocobalamin-transcobalamin II with an association constant (Ka) of 1.3 x 10(10) M-1 and 700 sites per cell. Binding was reversible, saturable and calcium dependent. Electron microscope radioautography following incubation with iodinated transcobalamin II at 4 degrees C led to a detectable labeling mainly restricted to the plasma membrane. 相似文献
77.
K. Ohta D. Keszenman-Pereyra T. Shibata A. Nicolas 《Molecular & general genetics : MGG》1996,250(4):395-404
Site-specific endonucleases have been found in various eukaryotic organelles such as mitochondria, chloroplasts and nuclei. These endonucleases initiate site-specific or homologous gene conversion in mitochondrial and nuclear DNA. Here, we report a new site-specific endonuclease activity, Endo.SK1, identified in mitochondria of strain SK1, a homothallic diploid strain ofSaccharomyces cerevisiae. Nucleotide sequences around the Endo.SK1-cleavage sites are different from those of known yeast site-specific endonucleases. The Endo.SK1 activity is, at least partly, specified by a gene in the SK1-derived mitochondria. A novel feature of the Endo.SK1 activity is its inducibility: the endonuclease activity was induced by ca. 40-fold by transfer of cells from a glucose medium into an acetate medium, and was then repressed. This transient induction was independent of the ploidy level of the cells, and coincided with induction of fumarase, a mitochondrial enzyme involved in the TCA cycle. Co-induction and co-repression of the mitochondrial site-specific endonuclease activity and a respiration-related enzyme indicate that the endonuclease activity is regulated in response to physiological conditions, and suggest a possible role for the endonuclease in mitochondrial DNA metabolism. 相似文献
78.
Identification of mutations in the ALD-gene of 20 families with adrenoleukodystrophy/adrenomyeloneuropathy 总被引:4,自引:0,他引:4
Ernst W. Krasemann V. Meier G. C. Korenke D. H. Hunneman F. Hanefeld 《Human genetics》1996,97(2):194-197
Adrenoleukodystrophy (ALD), an X-linked inherited metabolic disorder, is the most frequent inborn peroxisomal disease. It leads to demyelination in the central and peripheral nervous system. Defective -oxidation of saturated very long chain fatty acids (VLCFAs; C22:0–C26:0) in peroxisomes has been shown to lead to an accumulation of VLCFAs in leukoid areas of the central nervous system, peripheral nerves, adrenal gland, and blood. The ALD gene has been recently identified and encodes a 745-amino-acid protein. We screened patients with adrenoleukodystrophy/adrenomyeloneuropathy (ALD/AMN) from 20 kindreds for mutations in the ALD gene. Eleven missense and two nonsense mutations, five deletions, and one insertion were detected by direct sequencing of eight reverse transcribed fragments of the ALD-gene mRNA. Four mutations could be shown to be de novo. All mutations could be confirmed in carriers by sequencing genomic DNA. No correlation between the type of mutation and the severity of the phenotype could be observed. The mutations were not detected in the ALD gene of 30 healthy persons. 相似文献
79.
J.Michael Conlon Nicolas Chartrel Jerome Leprince Charles Suaudeau Jean Costentin Hubert Vaudry 《Peptides》1996,17(8):1291-1296
A peptide derived from the posttranslational processing of proenkephalin A was isolated from an extract of the brain of the European green frog Rana ridibunda and its primary structure established as: Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-Gly-Arg10-Pro-Glu-Trp-Trp-Gln-Asp-Tyr-Gln-Lys-Arg20-Tyr-Gly-Gly-Phe-Met. The structure was confirmed by chemical synthesis. The peptide represents an amphibian equivalent of bovine adrenal peptide E [preproenkephalin A (206–230)-peptide] but the sequence contains two amino acid substitutions (Met15 → Gln and Leu25 → Met) compared with the mammalian peptide. The data support previous hypotheses that the Leu-enkephalin sequence is not present in preproenkephalin A of amphibians. Intracerebroventricular injections of frog peptide E (10 and 100 ng) in mice had no significant effect on horizontal locomotor activity. The peptide, in doses up to 1 μg, had no effect on latency of escape jumping in the hot plate test and the peptide (100 ng) did not modify responses (paw licking, rearing, and escape jumping) in morphine-treated mice. 相似文献
80.
pHS-2 is a 3-kb plasmid originally isolated fromShigella flexneriinfections associated with reactive arthritis in humans. This plasmid is stably maintained in many clinical isolates ofShigella flexneri.The nucleotide sequence of this plasmid displays two closely linked regions that may play a role in the maintenance of this plasmid. One region consists of a 250-bp locus showing a significant homology to the ColE1cersite. The results indicate that thecer-like site of pHS-2, like the ColE1cersite, acts as arecA-independent, site-specific recombination site involved in the resolution of multimers, requiring the presence of the host-encoded factors ArgR, PepA, XerC, and XerD. The second region consists of a 36-kDa open reading frame involved in generating resistance to the bactericidal effect of complement, which confers a selective advantage to cells containing this sequence. The results also indicate that pHS-2 can replicate in another species of Enterobacteriaceae (Escherichia coli) and is mobilized by the F plasmid. 相似文献