全文获取类型
收费全文 | 113140篇 |
免费 | 1758篇 |
国内免费 | 893篇 |
专业分类
115791篇 |
出版年
2024年 | 27篇 |
2023年 | 66篇 |
2022年 | 243篇 |
2021年 | 414篇 |
2020年 | 247篇 |
2019年 | 311篇 |
2018年 | 12217篇 |
2017年 | 10919篇 |
2016年 | 8017篇 |
2015年 | 1691篇 |
2014年 | 1500篇 |
2013年 | 1674篇 |
2012年 | 5970篇 |
2011年 | 14320篇 |
2010年 | 12934篇 |
2009年 | 9009篇 |
2008年 | 10998篇 |
2007年 | 12415篇 |
2006年 | 1298篇 |
2005年 | 1465篇 |
2004年 | 1909篇 |
2003年 | 1780篇 |
2002年 | 1549篇 |
2001年 | 880篇 |
2000年 | 792篇 |
1999年 | 448篇 |
1998年 | 175篇 |
1997年 | 151篇 |
1996年 | 128篇 |
1995年 | 87篇 |
1994年 | 89篇 |
1993年 | 98篇 |
1992年 | 177篇 |
1991年 | 162篇 |
1990年 | 95篇 |
1989年 | 110篇 |
1988年 | 86篇 |
1987年 | 78篇 |
1986年 | 70篇 |
1985年 | 53篇 |
1984年 | 54篇 |
1983年 | 54篇 |
1982年 | 31篇 |
1978年 | 29篇 |
1976年 | 32篇 |
1975年 | 34篇 |
1973年 | 33篇 |
1972年 | 265篇 |
1971年 | 297篇 |
1970年 | 27篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
161.
ATP binding cassette transport systems account for most import of necessary nutrients in bacteria. The periplasmic binding component (or an equivalent membrane-anchored protein) is critical to recognizing cognate ligand and directing it to the appropriate membrane permease. Here we report the X-ray structures of d-xylose binding protein from Escherichia coli in ligand-free open form, ligand-bound open form, and ligand-bound closed form at 2.15 Å, 2.2 Å, and 2.2 Å resolutions, respectively. The ligand-bound open form is the first such structure to be reported at high resolution; the combination of the three different forms from the same protein furthermore gives unprecedented details concerning the conformational changes involved in binding protein function. As is typical of the structural family, the protein has two similar globular domains, which are connected by a three-stranded hinge region. The open liganded structure shows that xylose binds first to the C-terminal domain, with only very small conformational changes resulting. After a 34° closing motion, additional interactions are formed with the N-terminal domain; changes in this domain are larger and serve to make the structure more ordered near the ligand. An analysis of the interactions suggests why xylose is the preferred ligand. Furthermore, a comparison with the most closely related proteins in the structural family shows that the conformational changes are distinct in each type of binding protein, which may have implications for how the individual proteins act in concert with their respective membrane permeases. 相似文献
162.
B Brocks P Garin-Chesa E Behrle J E Park W J Rettig K Pfizenmaier D Moosmayer 《Molecular medicine (Cambridge, Mass.)》2001,7(7):461-469
BACKGROUND: Fibroblast activation protein (FAP) is a type II membrane protein expressed on tumor stroma fibroblasts in more than 90% of all carcinomas. FAP serves as a diagnostic marker and is potential therapeutic target for treatment of a wide variety of FAP+ carcinomas. Murine tumor stroma models and FAP-specific antibodies are required to investigate the functional role of FAP in tumor biology and its usefulness for drug targeting. We here describe the development of antibodies with crossreactivity for human (hFAP) and murine FAP (mFAP), which share 89% amino acid identity. MATERIAL AND METHODS: An FAP-/- mouse was sequentially immunized with recombinant murine and human FAP-CD8 fusion proteins. Immunoglobulin cDNA derived from hyperimmune spleen cells was used for the construction of a combinatorial single chain Fv (scFv) library. Phage display selection of FAP-specific scFv was performed on immobilized hFAP followed by selection on cells expressing murine FAP. RESULTS: High-affinity, species-crossreactive, FAP-specific scFv were isolated upon sequential phage display selection. A bivalent derivative (minibody M036) constructed thereof was applied for immunohistochemical analyses and allowed detection of FAP expression on stroma cells of different human carcinomas as well as on murine host stroma in a tumor xenograft model. CONCLUSIONS: MB M036, derived from phage display selected species crossreactive scFv, is suitable for tumor stroma targeting and will be a valuable tool in the analyses of the functional role of FAP in tumor biology as well as in the evaluation of the suitability of FAP for drug targeting. 相似文献
163.
Don A. Driscoll Annabel L. Smith Samantha Blight John Maindonald 《Biodiversity and Conservation》2012,21(6):1607-1625
Altered fire regimes are a driver of biodiversity decline. To plan effective management, we need to know how species are influenced
by fire and to develop theory describing fire responses. Animal responses to fire are usually measured using methods that
rely on animal activity, but animal activity may vary with time since fire, potentially biasing results. Using a novel approach
for detecting bias in the pit-fall trap method, we found that leaf-litter dependent reptiles were more active up to 6 weeks
after fire, giving a misleading impression of abundance. This effect was not discovered when modelling detectability with
zero-inflated binomial models. Two species without detection bias showed early-successional responses to time since fire,
consistent with a habitat-accommodation succession model. However, a habitat specialist did not have the predicted low abundance
after fire due to increased post-fire movement and non-linear recovery of a key habitat component. Interactions between fire
and other processes therefore must be better understood to predict reptile responses to changing fire-regimes. We conclude
that there is substantial bias when trapping reptiles after fire, with species that are otherwise hard to detect appearing
to be abundant. Studies that use a survey method based on animal activity such as bird calls or animal movements, likely face
a similar risk of bias when comparing recently-disturbed with control sites. 相似文献
164.
Little information exists on mixed-species groups between primates and other mammals in Neotropical forests. In this paper, we describe three such associations observed during an extensive large-vertebrate survey in central Amazonia, Brazil. Mixed-species groups between a primate species and another mammal were observed on seven occasions between squirrel monkeys (Saimiri cf. ustus) and either South American coatis (Nasua nasua) or tayras (Eira barbara) and between brown capuchins (Cebus apella) and coatis. All associations were restricted to floodplain forest during its dry stage. We suggest that the associations involving the coatis are connected to foraging and vigilance but may be induced by a common alternative food resource at a time of food shortage. 相似文献
165.
Lentiviral vectors have been used for gene transfer into the liver but their ability to efficiently transduce quiescent hepatocytes
remains controversial. Lentivirus-mediated gene transfer is more efficient in cycling cells. We determine the effect of H-IL6
in the lentiviral transduction. The lentiviral vector was used to transduce HepG2 cells and mice liver cells, previously treated
with H-IL6. The highest transduction level was observed in HepG2 cells treated with 30 ng/mL H-IL6 and in the mice that received
4 μg H-IL6. Our results suggest that H-IL6 is an inducer of lentiviral gene transfer into the liver cells without any toxicity. 相似文献
166.
Thomas Proft 《Biotechnology letters》2010,32(1):1-10
Sortases are transpeptidases produced by Gram-positive bacteria to anchor cell surface proteins covalently to the cell wall.
The Staphylococcus aureus sortase A (SrtA) cleaves a short C-terminal recognition motif (LPXTG) on the target protein followed by the formation of an amide bond with the pentaglycine
cross-bridge in the cell wall. Over recent years, several researchers have exploited this specific reaction for a range of
biotechnology applications, including the incorporation of non-native peptides and non-peptidic molecules into proteins, the
generation of nucleic acid–peptide conjugates and neoglycoconjugates, protein circularisation, and labelling of cell surface
proteins on living cells. 相似文献
167.
Seedlings of two barley genotypes (‘Maresi’ and wild form of Hordeum spontaneum) were treated with jasmonic acid (JA 5 μM and 15 μM) for 24 h, and then subjected to water stress (PEG 6000 solution of −
1.5 MPa). JA caused an increase in the content of ABA but not in that of proline and spermidine in the two studied genotypes.
The effect of the treatment did not depend on the applied JA concentration. The pre-stress treatment with JA changed plant
response to water deficit with regard to membrane injury. Treatment with a lower JA concentration (5 μM) caused a substantial
reduction of the stress-induced membrane damage in the both genotypes. A higher JA concentration (15 μM) caused the reduction
of membrane injury only in H. spontaneum and was ineffective in ‘Maresi’. JA had no influence on the leaf water status in water-stressed plants. A possible role of
JA in leaf ABA accumulation and alleviation of cell membrane injury under water deficit is discussed.
The work was partly supported by the Polish Committee For Scientific Research, grant No 5 PO6A 036 18 相似文献
168.
Aims
The present study was planned to investigate the diversity of 1-aminocyclopropane-1-carboxylate (ACC) deaminase producing bacteria from the rhizosphere of wheat plants and subsequent evaluation of selected PGPR on growth enhancement of wheat seedlings under drought and saline conditions.Methods
ACC deaminase producing plant growth promoting rhizobacteria (PGPR) were isolated from the rhizosphere of wheat and identified using 16S rRNA gene sequence analysis. Isolates were evaluated for various direct and indirect plant growth promoting (PGP) traits. Plant inoculation experiment was conducted using isolates IG 19 and IG 22 in wheat to assess their plant growth promotion potential under salinity and drought stress.Results
Thirty-eight ACC deaminase producing PGPR were isolated which belonged to 12 distinct genera and falling into four phyla γ-proteobacteria, β-proteobacteria, Flavobacteria and Firmicutes. Klebsiella sp. was the most abundant genera and followed by Enterobacter sp. The isolates exhibited ACC deaminase activities ranging from 0.106–0.980 μM α- ketobutyrate μg protein?1 h?1. The isolates showed multiple PGP traits such as IAA production, phosphate, zinc, potassium solubilization and siderophore production. Enterobacter cloacae (IG 19) and Citrobacter sp. (IG 22) inoculated wheat seedlings showed notable increases in fresh and dry biomass under non-stress as well as under stressed condition.Conclusion
To the best of our knowledge this is the first report of presence of ACC deaminase activity and other PGP traits from the genus Citrobacter and Empedobacter. Our finding revealed that the γ-proteobacteria group dominated the wheat rhizosphere. Plant inoculation with PGPR could be a sustainable approach to alleviate abiotic stresses in wheat plants. These native PGPR isolates could be used as potential biofertilizers for sustainable agriculture.169.
Hiroyuki Inoue Chiaki Kitao Shinichi Yano Shigeki Sawayama 《World journal of microbiology & biotechnology》2016,32(11):186
On-site cellulase and hemicellulase production is a promising way to reduce enzyme cost in the commercialization of the lignocellulose-to-ethanol process. A hemicellulase-producing fungal strain suitable for on-site enzyme production was selected from cultures prepared using wet disc-milling rice straw (WDM-RS) and identified as Trichoderma asperellum KIF125. KIF125 hemicellulase showed uniquely high abundance of β-xylosidase in the xylanolytic enzyme system compared to other fungal hemicellulase preparations. Supplementation of Talaromyces cellulolyticus cellulase with KIF125 hemicellulase was more effective than that with the hemicellulases from other fungal sources in reducing the total enzyme loading for the improvement of xylose yield in the hydrolysis of ball-milling RS, due to its high β-xylosidase dominance. β-Xylosidase in KIF125 hemicellulase was purified and classified as a glycosyl hydrolase family 3 enzyme with relatively high specificity for xylobiose. The production of KIF125 β-xylosidase in the fermentor was estimated as 118 U/g-WDM-RS (2350 U/L culture) at 48 h. These results demonstrate that KIF125 is promising as a practical hemicellulase source to combine with on-site cellulase production using T. cellulolyticus. 相似文献
170.
MicroRNA biogenesis and function in higher plants 总被引:1,自引:0,他引:1