全文获取类型
收费全文 | 4420篇 |
免费 | 303篇 |
出版年
2023年 | 28篇 |
2022年 | 53篇 |
2021年 | 96篇 |
2020年 | 96篇 |
2019年 | 111篇 |
2018年 | 119篇 |
2017年 | 107篇 |
2016年 | 153篇 |
2015年 | 251篇 |
2014年 | 237篇 |
2013年 | 276篇 |
2012年 | 322篇 |
2011年 | 306篇 |
2010年 | 196篇 |
2009年 | 174篇 |
2008年 | 233篇 |
2007年 | 205篇 |
2006年 | 196篇 |
2005年 | 174篇 |
2004年 | 190篇 |
2003年 | 155篇 |
2002年 | 121篇 |
2001年 | 66篇 |
2000年 | 54篇 |
1999年 | 56篇 |
1998年 | 34篇 |
1997年 | 24篇 |
1996年 | 32篇 |
1995年 | 25篇 |
1994年 | 24篇 |
1992年 | 32篇 |
1991年 | 34篇 |
1990年 | 21篇 |
1989年 | 36篇 |
1988年 | 26篇 |
1987年 | 19篇 |
1986年 | 20篇 |
1985年 | 19篇 |
1984年 | 34篇 |
1983年 | 22篇 |
1982年 | 33篇 |
1981年 | 16篇 |
1979年 | 23篇 |
1978年 | 14篇 |
1976年 | 19篇 |
1975年 | 18篇 |
1974年 | 18篇 |
1973年 | 20篇 |
1972年 | 17篇 |
1969年 | 19篇 |
排序方式: 共有4723条查询结果,搜索用时 187 毫秒
51.
I. G. Macreadie L. A. Castelli A. C. Ward M. J. R. Stark A. A. Azad 《Biotechnology letters》1993,15(3):213-218
Summary A convenient shuttle vector that enables high level secretion of proteins from Kluyveromyces lactis has been developed. The vector, pEPS1, contains a unique cloning site that allows the construction, in a single ligation step, of episomal plasmids capable of directing secretion of foreign gene products from K. lactis. As an example we demonstrate the production of -lactamase and determine optimal conditions for its secretion into the culture media. 相似文献
52.
Paula Tatiana Lopes Seixas Antonio Jacinto Demuner Luiz Claudio Almeida Barbosa Cristiane Isaac Cerceau Daiane Einhardt Blank Marcelo Henrique Dos Santos Elizeu de Sá Farias Marcelo Coutinho Picanço 《Journal of Applied Entomology》2023,147(2):116-125
Aedes aegypti L. (Diptera: Culicidae) is a vector for serious diseases in tropical regions. This pest is mainly controlled by commercial larvicides but the application of such products has led to environmental problems. Essential oils (EO) have been consistently reported as molecules with insecticidal activity and can be used to produce more environmentally friendly larvicides in the control of A. aegypti. In this study, the larvicidal effect of essential oils (EO) from the leaves of three Artemisia species was evaluated against A. aegypti. The oils were obtained from steam distillation and their chemical composition was determined by gas chromatography–mass spectrometry. The EO of Artemisia camphorata was the most active in the screening bioassay and presented LC50 and LC95 of 64.95 and 74.18 μg ml−1, respectively. In addition, we found that germacrene D-4-ol was the constituent responsible for the toxicity of this EO. Artemisia camphorata EO and its major constituent, germacrene D-4-ol, are promising for the development of natural larvicides against A. aegypti. 相似文献
53.
Roland E. Stark 《Ethology : formerly Zeitschrift fur Tierpsychologie》1992,91(4):301-310
Whether an individual strives to breed or restrains from reproducing directly but increases the fitness of another individual through its help, may be viewed as a result of a trade-off between fitness costs and benefits arising from this decision. A population of the large carpenter bee Xylocopa sulcatipes Maa was studied in southeast Israel over a two year period. Female reproductive success, in terms of genetic representation in the gene pool (genetic gain), was calculated using coefficients of relatedness and the number of offspring produced by solitary, reproductively dominant and helper bees of social nests. In one year helper females, that shared a nest with a related female accrued a higher genetic gain than did solitary females. In the subsequent year solitary females did better than the helper bees. In nests founded by unrelated females a helper bee accrued only little genetic gain. However, evidence is presented to show that subordinate behaviour can have an adaptive value and can ultimately be beneficial through nest inheritance by the subordinate bee. 相似文献
54.
Epoxide hydrolase (EC 3.3.2.3) activity was measured with [1-14C]cis-9,10-epoxystearic acid as the substrate. Homogenates were prepared from the endosperm tissue of germinating seeds of castor bean (Ricinus communis L. zanzibariensis). The activity of fatty-acid epoxide hydrolase was characterized with respect to dependence on time, amount of protein, pH and temperature. Analyses of enzyme distribution in endosperm, cotyledons, root and hypocotyl showed the highest total activity in the endosperm, less in the cotyledons and low activity in the root and hypocotyl. The specific activity was similar for cotyledons and endosperm. Analysis of the temporal expression of the enzyme in the endosperm during germination revealed high activity already in the imbibed seed. Activity was maximal between days four to six and then decreased at the end of one week. Subcellular fractionation of endosperm revealed a dual distribution of activity between the glyoxysomal and the cytosolic fractions. 相似文献
55.
56.
The gene (vgb) encoding the hemoglobin (VtHb) of Vitreoscilla sp. was cloned into a broad-host-range vector and stably transformed into Pseudomonas putida, Pseudomonas aeruginosa, and Xanthomonas maltophilia. vgb was stably maintained and expressed in functional form in all three species. When growth of the P. aeruginosa and X. maltophilia transformants in Luria-Bertani medium was compared with that of each corresponding untransformed strain, the VtHb-producing strains reached slightly higher maximum viable cell numbers, had significantly increased viability after extebded times in culture, and, like E. coli that produces VtHb, had significantly lower respiration rates. The VtHb-producing strain of P. putida also reached a slightly higher maximum viable cell number than its corresponding untransformed strain, but was significantly less viable after extended times in culture and, unlike the case in E. coli, had a generally higher respiration rate than the untransformed strain. When growth was monitored by absorbance, the results were similar to those obtained with viable cell counts. 相似文献
57.
Tn552 transposase purification and in vitro activities. 总被引:1,自引:0,他引:1
The Staphylococcus aureus transposon Tn552 encodes a protein (p480) containing the 'D,D(35)E' motif common to retroviral integrases and the transposases of a number of bacterial elements, including phage Mu, the integron-containing element Tn5090, Tn7 and IS3. p480 and a histidine-tagged derivative were overexpressed in Escherichia coli and purified by methods involving denaturation and renaturation. DNase I footprinting and gel binding assays demonstrated that p480 binds to two adjacent, directly repeated 23 bp motifs at each end of Tn552. Although donor strand cleavage by p480 was not detected, in vitro conditions were defined for strand transfer activity with transposon end fragments having pre-cleaved 3' termini. Strand transfer was Mn(2+)-dependent and appeared to join a single left or right end fragment to target DNA. The importance of the terminal dinucleotide CA-3' was demonstrated by mutation. The in vitro activities of p480 are consistent with its proposed function as the Tn552 transposase. 相似文献
58.
The Saccharomyces cerevisiae gene SDS22 encodes a potential regulator of the mitotic function of yeast type 1 protein phosphatase. 总被引:8,自引:2,他引:6 下载免费PDF全文
In higher eukaryotes, the activity and specificity of the type 1 protein serine-threonine phosphatase (PP1) catalytic subunit is thought to be controlled by its association with a number of regulatory or targeting subunits. Here we describe the characterization of a gene encoding one such potential polypeptide in the yeast Saccharomyces cerevisiae. The gene which we have isolated (termed SDS22) encodes a product with a high degree of sequence identity to the fission yeast sds22 protein, a known regulator of the mitotic function of PP1 in Schizosaccharomyces pombe. Using two different criteria, we have demonstrated that Sds22p and the catalytic subunit of PP1 (Glc7p) interact in yeast cells. We have also generated a temperature-sensitive allele of GLC7 (glc7-12) which causes a block to the completion of mitosis at the restrictive temperature. Additional copies of SDS22 lead to allele-specific suppression of the glc7-12 mutant, strongly suggesting that the interaction between the two proteins is of functional significance. Sds22p is therefore likely to be the second example of a PP1 regulatory subunit identified in S. cerevisiae. 相似文献
59.
Barlette Vania Elisabeth Garbujo Fábio Luiz Laurenti Freitas Luiz Carlos Gomide 《Molecular Engineering》1997,7(3-4):439-455
A five site potential model combining Lennard–Jones plus Coulomb potential functions has been developed for chloroform molecule.
The partial charges needed for Coulombic interactions were derived using the chelpg procedure implemented in the gaussian
92 program. These calculations were performed at the MP2 level with MC-311G* basis set for Cl and 6-311G** for C and H atoms.
The parameters for the Lennard–Jones potentials were optimized to reproduce experimental values for the density and enthalpy
of vaporization of the pure liquid at 298 K and 1 atm. The statistical mechanics calculations were performed with the Monte
Carlo method in the isothermic and isobaric (NpT) ensemble. Besides the values obtained for density, ρ, and molar enthalpy
of vaporization at constant pressure, Δ HV, for liquid chloroform, results for molar volume, Vm, molar heat capacity, Cp, isobaric thermal expansivity, αp, and isothermal compressibility, κT, for this pure liquid are also in very good agreement with experimental observations. Size effects on the values of thermodynamic
properties were investigated. The potential model was also tested by computing the free energy for solvating one chloroform
molecule into its own liquid at 298 K using a statistical perturbation approach. The result obtained compares well with the
experimental value. Site–site pair correlation functions were calculated and are in good accordance with theoretical results
available in the literature. Dipole–dipole correlation functions for the present five site model were also calculated at different
carbon–carbon distances. These correlations were compared to those obtained using the four site model reported in the literature.
An investigation of the solvent dependence of the relative free energy for cis/trans conversion of a hypothetical solute in
TIP4P water and chloroform was accomplished. The results show strong interaction of water and chloroform molecules with the
gauche conformer. The value obtained for the free energy barrier for cis/trans rotation in TIP4P water is higher than that
for chloroform. This result is in agreement with the continuous theory for solvation as the conformer with higher dipole moment
is more favoured by the solvent with higher dieletric constant. The results also show an increase in entropy as the solute
goes from the cis to the trans geometry and this result is more appreciable in the aqueous solution.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
60.
Mutations in the Saccharomyces Cerevisiae Type 2a Protein Phosphatase Catalytic Subunit Reveal Roles in Cell Wall Integrity, Actin Cytoskeleton Organization and Mitosis 总被引:3,自引:0,他引:3 下载免费PDF全文
Temperature-sensitive mutations were generated in the Saccharomyces cerevisiae PPH22 gene that, together with its homologue PPH21, encode the catalytic subunit of type 2A protein phosphatase (PP2A). At the restrictive temperature (37°), cells dependent solely on pph22(ts) alleles for PP2A function displayed a rapid arrest of proliferation. Ts(-) pph22 mutant cells underwent lysis at 37°, showing an accompanying viability loss that was suppressed by inclusion of 1 M sorbitol in the growth medium. Ts(-) pph22 mutant cells also displayed defects in bud morphogenesis and polarization of the cortical actin cytoskeleton at 37°. PP2A is therefore required for maintenance of cell integrity and polarized growth. On transfer from 24° to 37°, Ts(-) pph22 mutant cells accumulated a 2N DNA content indicating a cell cycle block before completion of mitosis. However, during prolonged incubation at 37°, many Ts(-) pph22 mutant cells progressed through an aberrant nuclear division and accumulated multiple nuclei. Ts(-) pph22 mutant cells also accumulated aberrant microtubule structures at 37°, while under semi-permissive conditions they were sensitive to the microtubule-destabilizing agent benomyl, suggesting that PP2A is required for normal microtubule function. Remarkably, the multiple defects of Ts(-) pph22 mutant cells were suppressed by a viable allele (SSD1-v1) of the polymorphic SSD1 gene. 相似文献