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931.
Rajiv Kumar Ranjan 《Analytical biochemistry》2010,402(1):91-92
Various methods of ligation are currently available and routinely used by molecular biologists, such as blunt end ligation, cohesive end (two and four overhangs), and ligation of Taq polymerase-derived products. However, there is no efficient method for the cloning of DNA fragments with 2-bp overhangs. We present a simple method for the efficient ligation of DNA fragments with 2-bp overhanging ends, ranging in size from 0.7 to 2.5 kbp. Our method involves the initial heating and flash freezing of the vector-insert DNA mix, and a subsequent unique ligation reaction. This method provides a new molecular biology tool for researchers. 相似文献
932.
Aggregation-prone Regions in HYPK Help It to Form Sequestration Complex for Toxic Protein Aggregates
Protein aggregates result from altered structural conformations and they can perturb cellular homeostasis. Prevention mechanisms, which function against protein aggregation by modulatory processes, are diverse and redundant. In this study, we have characterized Huntingtin interacting protein K (HYPK) as a global aggregation-regulatory protein. We report the mechanistic details of how HYPK's aggregation-prone regions allow it to sense and prevent other toxic protein's aggregation by forming unique annular-shaped sequestration complexes. Screenings for interacting partners of different aggregation-prone proteins identify HYPK as a global interacting partner/regulator of Huntingtin97Qexon1, α-Synuclein-A53T and Superoxide dismutase1-G93A. C-terminal hydrophobic region in HYPK makes direct contacts with aggregates to initiate the formation of sequestration complexes. HYPK acts as aggregate sensor by existing in a seeded amyloid-like state which also favors its own concentration-dependent self-oligomerization. Oligomerization of HYPK leads to annular and non-fibrillar/amorphous aggregates. Two hydrophobic segments in the C-terminus of HYPK are responsible for its own aggregations. Self-association of HYPK follows seed nucleation, in which oligomeric HYPK seeds nucleate to annular structures. Annular oligomers of HYPK fuse with each other to form amorphous aggregates. HYPK shows differential interactions with aggregation-prone and non-aggregating proteins, as it preferentially binds to aggregation-prone proteins with higher affinity than native/non-aggregating proteins. This favors the formation of HYPK's sequestration complexes both in cytosol and in ribosome. Besides having aggregation-preventive property, HYPK also reduces the cellular level of toxic proteins. In vivo, HYPK sequestration complexes prevent the formation of toxic protein aggregates to physiologically show positive impact on cell survival and restoration of normal cell physiology. 相似文献
933.
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935.
Sanghamitra Samantaray Gyana Ranjan Rout Premananda Das 《Acta Physiologiae Plantarum》1999,21(1):27-35
Callus of Trema orientalis derived from both contaminated and uncontaminated sources were tested in vitro for their relative tolerance to chromium and nickel. The calluses derived from contaminated source were metal-tolerant and
showed better growth than those obtained from uncontaminated plants. The specificity of metal tolerance shown by the parent
material was maintained in the calluses. Compared to the uncontaminated explants, the calli derived from contaminated sources
exhibited higher catalase and peroxidase activities but a reduced acid phosphatase activity. Biochemical studies, provided
evidence that the contaminated sources were physiologically distinct from the uncontaminated ones. Thus, this study indicated
that seeds of Trema orientalis collected from contaminated sites were tolerant to chromium and nickel, and may have the advantage of being used in sustainable
revegetation programmes on chromiferous minewastes. 相似文献
936.
The effect of aqueous leaf extracts of four plants, Argemone mexicana, Cyperus rotundus, Euphorbia hirta and Solanum nigrum , on growth and aflatoxin production by Aspergillus flavus was studied in SMKY liquid medium. All the plants inhibited aflatoxin production. No correlation between the growth of the fungus and aflatoxin synthesis was observed. The influence of these plants on the ratio of aflatoxin B1 to G1 is discussed. 相似文献
937.
Ranjan Banerjee Wimal Pathmasiri Rodney Snyder Susan McRitchie Susan Sumner 《Metabolomics : Official journal of the Metabolomic Society》2012,8(6):1012-1025
Phthalates are plasticizers finding wide spread use in industrial and household products, with measureable levels of phthalate-derived metabolites in the general US population. Phthalates have endocrine disruption potential and have been implicated as obesogens. Our exploratory investigation to reveal the impact of in utero exposure to a phthalate on the biochemical profiles of the brain, testes, and uterus of prepubertal offspring, and of tissues from dams administered butylbenzyl phthalate (BBP). Pregnant rats (three per group) were administered (on gestation day 14?C21) corn oil (control), or 25?mg/kg/day or 750?mg/kg/day BBP in corn oil. Tissues were collected from each of the dams on postnatal day (pnd) 21 (~3?weeks after the end of BBP administration), and from each of the pups on pnd 26 (~4 weeks after birth to dams administered vehicle or BBP during gestation) and processed for metabolomics analysis. Multivariate data analyses revealed metabolites that best distinguished the exposed and control groups. The metabolites most important to distinguishing the study groups were tested for significance using the exact Wilcoxon rank-sum test. Male pups had significant differences (control versus BBP dose groups) in levels of metabolites for both the brain and testes even at the P?<?0.01 level. However, female pups and dams had significant testing for the uterus only at the P?=?0.1 level tested. Female pups also had some significant differences for the brain with P values between 0.5 and 0.1. Amino acid metabolism (male and female pups) and phospholipid metabolism (male pups) were perturbed for the brain. Amino acid metabolism, purine metabolism, and TCA cycle were perturbed for tests and uterus. This study demonstrated the use of metabolomics to reveal metabolic perturbations in tissues of offspring following in utero exposures, and suggests the use of this approach for determining the impact of exposure past the time of the presence of the parent compound and metabolites derived from the parent compound. 相似文献
938.
Both sodium orthovanadate and fluoroaluminate were found to stimulate in vitro ovulation in intact goldfish follicles, suggesting the involvement of G-proteins in ovulation. Although orthovanadate was able to stimulate cAMP production, it probably stimulates ovulation by some other mechanism since cAMP blocks ovulation in this species. These agents also stimulated the accumulation of labeled inositol phosphate in follicle walls. The time course of inositol phosphate production showed a slightly delayed and continuous accumulation for isomers of inositol mono-, bis-, and trisphosphates. No change was observed in inositol tetrakisphosphate levels over time. The accumulation of inositol phosphates in response to orthovanadate was also dose-dependent. Lithium chloride (10 mM) caused varying increases in the levels of most isomers and a decrease in ins-3,4-P2. Inositol phosphate production varied significantly with changes in the maturational stage of follicles. Peak production was observed in follicles 7-8 h after hCG treatment, which corresponds almost exactly with the time of ovulation. This correlation of maximal inositol phosphate production with the time of ovulation, along with the stimulation of ovulation by diacylglycerols, a phorbol ester, and the G-protein-stimulating agents, orthovanadate and fluoroaluminate, suggests a role for polyphosphatidylinositol hydrolysis in ovulation. 相似文献
939.
Ranjan K. Pradhan 《Biophysical journal》2010,98(2):218-230
Sodium-calcium antiporter is the primary efflux pathway for Ca2+ in respiring mitochondria, and hence plays an important role in mitochondrial Ca2+ homeostasis. Although experimental data on the kinetics of Na+-Ca2+ antiporter are available, the structure and composition of its functional unit and kinetic mechanisms associated with the Na+-Ca2+ exchange (including the stoichiometry) remains unclear. To gain a quantitative understanding of mitochondrial Ca2+ homeostasis, a biophysical model of Na+-Ca2+ antiporter is introduced that is thermodynamically balanced and satisfactorily describes a number of independent data sets under a variety of experimental conditions. The model is based on a multistate catalytic binding mechanism for carrier-mediated facilitated transport and Eyring's free energy barrier theory for interconversion and electrodiffusion. The model predicts the activating effect of membrane potential on the antiporter function for a 3Na+:1Ca2+ electrogenic exchange as well as the inhibitory effects of both high and low pH seen experimentally. The model is useful for further development of mechanistic integrated models of mitochondrial Ca2+ handling and bioenergetics to understand the mechanisms by which Ca2+ plays a role in mitochondrial signaling pathways and energy metabolism. 相似文献
940.
Ranjan K Sahoo Mohammad W Ansari Madhusmita Pradhan Tushar K Dangar Santanu Mohanty Narendra Tuteja 《Protoplasma》2014,251(4):943-953
Beneficial microorganisms have been considered as an important tool for crop improvement. Native isolates of Azospirillum spp. were obtained from the rhizospheres of different rice fields. Phenotypic, biochemical and molecular characterizations of these isolates led to the identification of six efficient strain of Azospirillum. PCR amplification of the nif genes (nifH, nifD and nifK) and protein profile of Azospirillum strains revealed inter-generic and inter-specific diversity among the strains. In vitro nitrogen fixation performance and the plant growth promotion activities, viz. siderophore, HCN, salicylic acid, IAA, GA, zeatin, ABA, NH3, phosphorus metabolism, ACC deaminase and iron tolerance were found to vary among the Azospirillum strains. The effect of Azospirillum formulations on growth of rice var. Khandagiri under field condition was evaluated, which revealed that the native formulation of Azospirillum of CRRI field (As6) was most effective to elevate endogenous nutrient content, and improved growth and better yield are the result. The 16S rRNA sequence revealed novelty of native Azospirillum lipoferum (As6) (JQ796078) in the NCBI database. 相似文献