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991.
Rational siRNA design for RNA interference 总被引:166,自引:0,他引:166
Reynolds A Leake D Boese Q Scaringe S Marshall WS Khvorova A 《Nature biotechnology》2004,22(3):326-330
Short-interfering RNAs suppress gene expression through a highly regulated enzyme-mediated process called RNA interference (RNAi). RNAi involves multiple RNA-protein interactions characterized by four major steps: assembly of siRNA with the RNA-induced silencing complex (RISC), activation of the RISC, target recognition and target cleavage. These interactions may bias strand selection during siRNA-RISC assembly and activation, and contribute to the overall efficiency of RNAi. To identify siRNA-specific features likely to contribute to efficient processing at each step, we performed a systematic analysis of 180 siRNAs targeting the mRNA of two genes. Eight characteristics associated with siRNA functionality were identified: low G/C content, a bias towards low internal stability at the sense strand 3'-terminus, lack of inverted repeats, and sense strand base preferences (positions 3, 10, 13 and 19). Further analyses revealed that application of an algorithm incorporating all eight criteria significantly improves potent siRNA selection. This highlights the utility of rational design for selecting potent siRNAs and facilitating functional gene knockdown studies. 相似文献
992.
Marco Fichera Angela Spalletta Flavia Fiorenza Turi Lombardo Gino Schilirò Ryad Tamouza Claudine Lapouméroulie Dominique Labie A. Ragusa 《Human genetics》1997,99(3):381-386
To evaluate the allelic frequency and genetic diversity of α-thalassemia defects in Sicily, both epidemiological and patient-oriented
studies were carried out. For the epidemiological study, phenotypic data were collected on more than 1000 Sicilian individuals.
Among them, 427 were explored at the molecular level for nine α-thalassemic variants known to be common in the Mediterranean
region. Our data reveal an allele frequency of 4.1% for α+-thalassemia matching that of β-thalassemia in this region. The presence of α°-thalassemia (––MEDI and ––CAL) was observed only in the group of referred patients. Newly acquired nucleotide sequence data on the deletional breakpoint
of ––CAL allowed us to design a simple PCR-based procedure for exploring this allele. The data also provide additional information
concerning the genetic mechanisms involved in such large deletions.
Received: 8 August 1996 / Revised: 16 October 1996 相似文献
993.
Stéphane Compant Marine C. Cambon Corinne Vacher Birgit Mitter Abdul Samad Angela Sessitsch 《Environmental microbiology》2021,23(4):1812-1829
The plant endosphere is colonized by complex microbial communities and microorganisms, which colonize the plant interior at least part of their lifetime and are termed endophytes. Their functions range from mutualism to pathogenicity. All plant organs and tissues are generally colonized by bacterial endophytes and their diversity and composition depend on the plant, the plant organ and its physiological conditions, the plant growth stage as well as on the environment. Plant-associated microorganisms, and in particular endophytes, have lately received high attention, because of the increasing awareness of the importance of host-associated microbiota for the functioning and performance of their host. Some endophyte functions are known from mostly lab assays, genome prediction and few metagenome analyses; however, we have limited understanding on in planta activities, particularly considering the diversity of micro-environments and the dynamics of conditions. In our review, we present recent findings on endosphere environments, their physiological conditions and endophyte colonization. Furthermore, we discuss microbial functions, the interaction between endophytes and plants as well as methodological limitations of endophyte research. We also provide an outlook on needs of future research to improve our understanding on the role of microbiota colonizing the endosphere on plant traits and ecosystem functioning. 相似文献
994.
Sara E. Cannon Simon D. Donner Angela Liu Pedro C. González Espinosa Andrew H. Baird Julia K. Baum Andrew G. Bauman Maria Beger Cassandra E. Benkwitt Matthew J. Birt Yannick Chancerelle Joshua E. Cinner Nicole L. Crane Vianney Denis Martial Depczynski Nur Fadli Douglas Fenner Christopher J. Fulton Yimnang Golbuu Nicholas A. J. Graham James Guest Hugo B. Harrison Jean-Paul A. Hobbs Andrew S. Hoey Thomas H. Holmes Peter Houk Fraser A. Januchowski-Hartley Jamaluddin Jompa Chao-Yang Kuo Gino Valentino Limmon Yuting V. Lin Timothy R. McClanahan Dominic Muenzel Michelle J. Paddack Serge Planes Morgan S. Pratchett Ben Radford James Davis Reimer Zoe T. Richards Claire L. Ross John Rulmal Jr. Brigitte Sommer Gareth J. Williams Shaun K. Wilson 《Global Change Biology》2023,29(12):3318-3330
Scientists and managers rely on indicator taxa such as coral and macroalgal cover to evaluate the effects of human disturbance on coral reefs, often assuming a universally positive relationship between local human disturbance and macroalgae. Despite evidence that macroalgae respond to local stressors in diverse ways, there have been few efforts to evaluate relationships between specific macroalgae taxa and local human-driven disturbance. Using genus-level monitoring data from 1205 sites in the Indian and Pacific Oceans, we assess whether macroalgae percent cover correlates with local human disturbance while accounting for factors that could obscure or confound relationships. Assessing macroalgae at genus level revealed that no genera were positively correlated with all human disturbance metrics. Instead, we found relationships between the division or genera of algae and specific human disturbances that were not detectable when pooling taxa into a single functional category, which is common to many analyses. The convention to use percent cover of macroalgae as an indication of local human disturbance therefore likely obscures signatures of local anthropogenic threats to reefs. Our limited understanding of relationships between human disturbance, macroalgae taxa, and their responses to human disturbances impedes the ability to diagnose and respond appropriately to these threats. 相似文献
995.
Ian P. Holmes Fabrizio Micheli Simon Gaines Olivier Lorthioir Steve P. Watson Romano Di Fabio Gabriella Gentile Christian Heidbreder Chiara Savoia Angela Worby 《Bioorganic & medicinal chemistry letters》2009,19(16):4799-4801
The synthesis and SAR of a new series of potent and selective dopamine D3 receptor antagonists is reported. 相似文献
996.
SUMMARY: Haplotypes carry important information that can direct investigators towards underlying susceptibility variants, and hence multiple tagging single nucleotide polymorphisms (tSNPs) are usually studied in candidate gene association studies. However, it is often unknown which SNPs should be included in haplotype analyses, or which tests should be performed for maximum power. We have developed a program, hapConstructor, which automatically builds multi-locus SNP sets to test for association in a case-control framework. The multi-SNP sets considered need not be contiguous; they are built based on significance. An important feature is that the missing data imputation is carried out based on the full data, for maximal information and consistency. HapConstructor is implemented in a Monte Carlo framework and naturally extends to allow for significance testing and false discovery rates that account for the construction process and to related individuals. HapConstructor is a useful tool for exploring multi-locus associations in candidate genes and regions. AVAILABILITY: http://www-genepi.med.utah.edu/Genie. 相似文献
997.
Vincenzo Guida Maria Cantarella Angela Chambery Maria C. Mezzacapo Augusto Parente Nicola Landi Valeria Severino Antimo Di Maro 《Molecular biotechnology》2014,56(8):738-746
Four novel basic peroxidases, named AaP-1, AaP-2, AaP-3, and AaP-4, were purified from Asparagus acutifolius L. seeds by cation-exchange and gel filtration chromatographies. The four proteins showed a similar electrophoretic mobility of 46 kDa while, by MALDI–TOF MS, different Mr values of 42758.3, 41586.9, 42796.3, and 41595.5 were determined for AaP-1, AaP-2, AaP-3, and AaP-4, respectively. N-terminal sequences of AaPs 1–4 up to residue 20 showed a high percentage of identity with the peroxidase from Glycine max. In addition, AaP-1, AaP-2, AaP-3, and AaP-4 were found to be glycoproteins, containing 21.75, 22.27, 25.62, and 18.31 % of carbohydrates, respectively. Peptide mapping and MALDI–TOF MS analysis of AaPs 1–4 showed that the structural differences between AaP-1 and AaP-2 and AaP-3 and AaPs-4 were mainly due to their glycan content. We also demonstrate that AaPs were able to remove phenolic compounds from olive oil mill wastewaters with a higher catalytic efficiency with respect to horseradish peroxidase, thus representing candidate enzymes for potential biotechnological applications in the environmental field. 相似文献
998.
999.
Luciana A. Haddad Regina C. Mingroni-Netto Angela M. Vianna-Morgante Sérgio D. J. Pena 《Human genetics》1996,97(6):808-812
Ever since the identification of the genetic cause of fragile X syndrome as the expansion of an unstable trinucleotide sequence,
several diagnostic strategies have evolved from molecular studies. However, we still lack a simple test suitable for population
screening. We have therefore developed a nonisotopic polymerase chain reaction (PCR)-based technique for the identification
of fragile X full mutations among men, with easy visualization of the PCR products on silver-stained polyacrylamide gels.
The technique consists of PCR amplification with primers that flank the trinucleotide repeats, with a product of 557 bp for
the (CGG)29 allele. Conditions were established such that full mutations failed to amplify and were thus identified with 98% sensitivity
compared with Southern blot analysis. To produce an indispensable internal control we added to the reaction a third primer,
internal to this fragment, allowing the multiplex amplification of a monomorphic band corresponding to a CG-rich stretch 147
bp upstream of the polymorphic region. In trials involving 41 patients and 74 controls, the PCR-based test here described
showed specificity of more than 98.6%, accuracy of 99% and a sensitivity of 98%. Thus, although not suitable for medical diagnosis,
it constitutes a useful tool for screening for the fragile X syndrome in populations of mentally retarded males.
Received: 31 May 1995 / Revised: 4 October 1995 相似文献
1000.
Ito M Nakajima S Fujisawa D Miyashita M Kim Y Shear MK Ghesquiere A Wall MM 《PloS one》2012,7(2):e31209