全文获取类型
收费全文 | 6298篇 |
免费 | 438篇 |
国内免费 | 1篇 |
出版年
2024年 | 3篇 |
2023年 | 23篇 |
2022年 | 64篇 |
2021年 | 135篇 |
2020年 | 97篇 |
2019年 | 149篇 |
2018年 | 193篇 |
2017年 | 138篇 |
2016年 | 248篇 |
2015年 | 355篇 |
2014年 | 410篇 |
2013年 | 485篇 |
2012年 | 634篇 |
2011年 | 535篇 |
2010年 | 390篇 |
2009年 | 321篇 |
2008年 | 450篇 |
2007年 | 424篇 |
2006年 | 340篇 |
2005年 | 297篇 |
2004年 | 311篇 |
2003年 | 244篇 |
2002年 | 184篇 |
2001年 | 50篇 |
2000年 | 44篇 |
1999年 | 47篇 |
1998年 | 41篇 |
1997年 | 18篇 |
1996年 | 11篇 |
1995年 | 10篇 |
1994年 | 6篇 |
1993年 | 8篇 |
1992年 | 12篇 |
1991年 | 8篇 |
1990年 | 4篇 |
1989年 | 4篇 |
1988年 | 2篇 |
1987年 | 3篇 |
1986年 | 5篇 |
1985年 | 3篇 |
1983年 | 3篇 |
1982年 | 4篇 |
1980年 | 2篇 |
1976年 | 2篇 |
1975年 | 2篇 |
1970年 | 2篇 |
1969年 | 2篇 |
1968年 | 2篇 |
1967年 | 3篇 |
1965年 | 2篇 |
排序方式: 共有6737条查询结果,搜索用时 375 毫秒
101.
Key message
Arabidopsis gulliver3 - D/dwarf4 - D displays growth-promoting phenotypes due to activation tagging of a key brassinosteroid biosynthetic gene DWARF4. In gul3-D/dwf4-D , the Jasmonate and Salicylate signaling pathways were relatively activated and suppressed, respectively.Abstract
Energy allocation between growth and defense is elegantly balanced to achieve optimal development in plants. Brassinosteroids (BRs), steroidal hormones essential for plant growth, are regulated by other plant hormones, including auxin and jasmonates (JA); auxin stimulates the expression of a key brassinosteroid (BR) biosynthetic gene, DWARF4 (DWF4), whereas JA represses it. To better understand the interaction mechanisms between growth and defense, we isolated a fast-growing mutant, gulliver3-D (gul3-D), that resulted from the activation tagging of DWF4, and examined the response of this mutant to defense signals, including JA, Pseudomonas syringae pv. tomato (Pst DC3000) infection, and wounding. The degree of root growth inhibition following MeJA treatment was significantly decreased in gul3-1D/dwf4-5D relative to the wild type, suggesting that JA signaling is partially desensitized in gul3-1D. Quantitative RT-PCR analysis of the genes involved in JA and salicylic acid (SA) responses, including MYC2, PDF1.2, CORI3, PR1, and PR2, revealed that JA signaling was preferentially activated in gul3-1D, whereas SA signaling was suppressed. As a result, gul3-1D was more susceptible to a biotrophic pathogen, Pst DC3000. Based on our results, we propose a model in which BR and JA cooperate to balance energy allocation between growth and defense responses. In ambient conditions, BRs promote plant growth; however, when stresses trigger JA signaling, JA compromises BR signaling by downregulating DWF4 expression. 相似文献102.
Ellen H. Esch Daniel L. Hernández Jae R. Pasari Rose S. G. Kantor Paul C. Selmants 《Plant and Soil》2013,366(1-2):671-682
Background and aims
Exotic species, nitrogen (N) deposition, and grazing are major drivers of change in grasslands. However little is known about the interactive effects of these factors on below-ground microbial communities.Methods
We simulated realistic N deposition increases with low-level fertilization and manipulated grazing with fencing in a split-plot experiment in California’s largest serpentine grassland. We also monitored grazing intensity using camera traps and measured total available N to assess grazing and nutrient enrichment effects on microbial extracellular enzyme activity (EEA), microbial N mineralization, and respiration rates in soil.Results
Continuous measures of grazing intensity and N availability showed that increased grazing and N were correlated with increased microbial activity and were stronger predictors than the categorical grazing and fertilization measures. Exotic cover was also generally correlated with increased microbial activity resulting from exotic-driven nutrient cycling alterations. Seasonal effects, on abiotic factors and plant phenology, were also an important factor in EEA with lower activity occurring at peak plant biomass.Conclusions
In combination with previous studies from this serpentine grassland, our results suggest that grazing intensity and soil N availability may affect the soil microbial community indirectly via effects on exotic cover and associated changes in nutrient cycling while grazing directly impacts soil community function. 相似文献103.
104.
Hyo Jae Kang Young Sik Park Chang-Hoon Lee Sang-Min Lee Jae-Joon Yim Chul-Gyu Yoo Young Whan Kim Sung Koo Han Jong-Yil Chai Jinwoo Lee 《The Korean journal of parasitology》2013,51(5):569-572
Dirofilariasis is a rare disease in humans. We report here a case of a 48-year-old male who was diagnosed with pulmonary dirofilariasis in Korea. On chest radiographs, a coin lesion of 1 cm in diameter was shown. Although it looked like a benign inflammatory nodule, malignancy could not be excluded. So, the nodule was resected by video-assisted thoracic surgery. Pathologically, chronic granulomatous inflammation composed of coagulation necrosis with rim of fibrous tissues and granulations was seen. In the center of the necrotic nodules, a degenerating parasitic organism was found. The parasite had prominent internal cuticular ridges and thick cuticle, a well-developed muscle layer, an intestinal tube, and uterine tubules. The parasite was diagnosed as an immature female worm of Dirofilaria immitis. This is the second reported case of human pulmonary dirofilariasis in Korea. 相似文献
105.
Chinese hamster ovary (CHO) cells, that are widely used for production of therapeutic proteins, are subjected to apoptosis and autophagy under the stresses induced by conditions such as nutrient deprivation, hyperosmolality and addition of sodium butyrate. To achieve a cost-effective level of production, it is important to extend the culture longevity. Until now, there have been numerous studies in which apoptosis of recombinant CHO (rCHO) cells was inhibited, resulting in enhanced production of therapeutic proteins. Recently, autophagy in rCHO cells has drawn attention because it can be genetically and chemically controlled to increase cell survival and productivity. Autophagy is a global catabolic process which involves multiple pathways and genes that regulate the lysosomal degradation of intracellular components. A simultaneous targeting of anti-apoptosis and pro-autophagy could lead to more efficient protection of cells from stressful culture conditions. In this regard, it is worthwhile to have a detailed understanding of the autophagic pathway, in order to select appropriate genes and chemical targets to manage autophagy in rCHO cells, and thus to enhance the production of therapeutic proteins. 相似文献
106.
Seung Hwan Lee Kyoung-Hee Kang Eun Young Kim Tong Un Chae Young Hoon Oh Soon Ho Hong Bong Keun Song Jonggeon Jegals Si Jae Park Sang Yup Lee 《Biotechnology letters》2013,35(10):1631-1637
We have previously analyzed the proteome of recombinant Escherichia coli producing poly(3-hydroxybutyrate) [P(3HB)] and revealed that the expression level of several enzymes in central metabolism are proportional to the amount of P(3HB) accumulated in the cells. Based on these results, the amplification effects of triosephosphate isomerase (TpiA) and fructose-bisphosphate aldolase (FbaA) on P(3HB) synthesis were examined in recombinant E. coli W3110, XL1-Blue, and W lacI mutant strains using glucose, sucrose and xylose as carbon sources. Amplification of TpiA and FbaA significantly increased the P(3HB) contents and concentrations in the three E. coli strains. TpiA amplification in E. coli XL1-Blue lacI increased P(3HB) from 0.4 to 1.6 to g/l from glucose. Thus amplification of glycolytic pathway enzymes is a good strategy for efficient production of P(3HB) by allowing increased glycolytic pathway flux to make more acetyl-CoA available for P(3HB) biosynthesis. 相似文献
107.
Hyuk Lee Si Jae Park Mee-Jung Han Gyeong Tae Eom Min-Jung Choi Seong Ho Kim Young Hoon Oh Bong Keun Song Seung Hwan Lee 《Biotechnology letters》2013,35(10):1677-1683
Microbial-surface display is the expression of proteins or peptides on the surface of cells by fusing an appropriate protein as an anchoring motif. Here, the outer membrane protein W (OmpW) was selected as a fusion partner for functional expression of Pseudomonas fluorescence SIK W1 lipase (TliA) on the cell-surface of Escherichia coli. Localization of the truncated OmpW-TliA fusion protein on the cell-surface was confirmed by immunoblotting and functional assay of lipase activity. Enantioselective hydrolysis of rac-phenylethyl butanoate by the displayed lipase resulted in optically active (R)-phenyl ethanol with 96 % enantiomeric excess and 44 % of conversion in 5 days. Thus, a small outer membrane protein OmpW, is a useful anchoring motif for displaying an active enzyme of ~50 kDa on the cell-surface and the surface-displayed lipase can be employed as an enantioselective biocatalyst in organic synthesis. 相似文献
108.
Yudai Nagata Jae Man Lee Hiroaki Mon Shigeo Imanishi Sun Mee Hong Shoji Komatsu Yuji Oshima Takahiro Kusakabe 《Biotechnology letters》2013,35(7):1009-1016
Glycoproteins have various biological functions including enzymatic activity, protein stability and others. Due to the presence of paucimannosidic N-linked glycans, recombinant proteins from an insect cell expression system may not be suitable for therapeutic use. Because baculovirus expression systems (BESs) are used to produce recombinant proteins, it is of interest to modify the endogenous N-glycosylation pathway in insects to mimic that of mammals. Using a soaking RNAi sensitive cell line, BmN4-SID1, has enabled us to suppress Bombyx mori FDL (BmFDL), an N-linked glycan-specific β-N-acetylglucosaminidase. Western blotting and MALDI-TOF MS demonstrated that the BmFDL depletion almost completely converted the paucimannosidic structures of the recombinant proteins produced by BES into a complex-type structure. This highly efficient, simple and low-cost method can be used for mass production of secretion proteins with complex-type N-linked glycans. 相似文献
109.
Jeong Myeong Kim Hyo Jung Lee Dae Sung Lee Che Ok Jeon 《Applied and environmental microbiology》2013,79(6):1969-1979
To characterize the denitrifying phosphorus (P) uptake properties of “Candidatus Accumulibacter phosphatis,” a sequencing batch reactor (SBR) was operated with acetate. The SBR operation was gradually acclimated from anaerobic-oxic (AO) to anaerobic-anoxic-oxic (A2O) conditions by stepwise increases of nitrate concentration and the anoxic time. The communities of “Ca. Accumulibacter” and associated bacteria at the initial (AO) and final (A2O) stages were compared using 16S rRNA and polyphosphate kinase genes and using fluorescence in situ hybridization (FISH). The acclimation process led to a clear shift in the relative abundances of recognized “Ca. Accumulibacter” subpopulations from clades IIA > IA > IIF to clades IIC > IA > IIF, as well as to increases in the abundance of other associated bacteria (Dechloromonas [from 1.2% to 19.2%] and “Candidatus Competibacter phosphatis” [from 16.4% to 20.0%]), while the overall “Ca. Accumulibacter” abundance decreased (from 55.1% to 29.2%). A series of batch experiments combined with FISH/microautoradiography (MAR) analyses was performed to characterize the denitrifying P uptake properties of the “Ca. Accumulibacter” clades. In FISH/MAR experiments using slightly diluted sludge (∼0.5 g/liter), all “Ca. Accumulibacter” clades successfully took up phosphorus in the presence of nitrate. However, the “Ca. Accumulibacter” clades showed no P uptake in the presence of nitrate when the sludge was highly diluted (∼0.005 g/liter); under these conditions, reduction of nitrate to nitrite did not occur, whereas P uptake by “Ca. Accumulibacter” clades occurred when nitrite was added. These results suggest that the “Ca. Accumulibacter” cells lack nitrate reduction capabilities and that P uptake by “Ca. Accumulibacter” is dependent upon nitrite generated by associated nitrate-reducing bacteria such as Dechloromonas and “Ca. Competibacter.” 相似文献
110.
Ramesh Prasad Pandey Sailesh Malla Dinesh Simkhada Byung-Gee Kim Jae Kyung Sohng 《Applied microbiology and biotechnology》2013,97(5):1889-1901
Quercetin, a flavonol aglycone, is one of the most abundant flavonoids with high medicinal value. The bioavailability and pharmacokinetic properties of quercetin are influenced by the type of sugars attached to the molecule. To efficiently diversify the therapeutic uses of quercetin, Escherichia coli was harnessed as a production factory by the installation of various plant and bacterial UDP-xylose sugar biosynthetic genes. The genes encoding for the UDP-xylose pathway enzymes phosphoglucomutase (nfa44530), glucose-1-phosphate uridylyltransferase (galU), UDP-glucose dehydrogenase (calS8), and UDP-glucuronic acid decarboxylase (calS9) were overexpressed in E. coli BL21 (DE3) along with a glycosyltransferase (arGt-3) from Arabidopsis thaliana. Furthermore, E. coli BL21(DE3)/?pgi, E. coli BL21(DE3)/?zwf, E. coli BL21(DE3)/?pgi?zwf, and E. coli BL21(DE3)/?pgi?zwf?ushA mutants carrying the aforementioned UDP-xylose sugar biosynthetic genes and glycosyltransferase and the galU-integrated E. coli BL21(DE3)/?pgi host harboring only calS8, calS9, and arGt-3 were constructed to enhance whole-cell bioconversion of exogeneously supplied quercetin into 3-O-xylosyl quercetin. Here, we report the highest production of 3-O-xylosyl quercetin with E. coli BL21 (DE3)/?pgi?zwf?ushA carrying UDP-xylose sugar biosynthetic genes and glycosyltransferase. The maximum concentration of 3-O-xylosyl quercetin achieved was 23.78 mg/L (54.75 μM), representing 54.75 % bioconversion, which was an ~4.8-fold higher bioconversion than that shown by E. coli BL21 (DE3) with the same set of genes when the reaction was carried out in 5-mL culture tubes with 100 μM quercetin under optimized conditions. Bioconversion was further improved by 98 % when the reaction was scaled up in a 3-L fermentor at 36 h. 相似文献