全文获取类型
收费全文 | 521篇 |
免费 | 63篇 |
出版年
2022年 | 4篇 |
2021年 | 7篇 |
2020年 | 6篇 |
2019年 | 4篇 |
2018年 | 14篇 |
2017年 | 8篇 |
2016年 | 22篇 |
2015年 | 30篇 |
2014年 | 27篇 |
2013年 | 33篇 |
2012年 | 54篇 |
2011年 | 37篇 |
2010年 | 24篇 |
2009年 | 14篇 |
2008年 | 31篇 |
2007年 | 24篇 |
2006年 | 30篇 |
2005年 | 26篇 |
2004年 | 26篇 |
2003年 | 23篇 |
2002年 | 24篇 |
2001年 | 2篇 |
2000年 | 3篇 |
1999年 | 6篇 |
1998年 | 7篇 |
1997年 | 10篇 |
1996年 | 7篇 |
1995年 | 4篇 |
1994年 | 10篇 |
1993年 | 7篇 |
1992年 | 5篇 |
1991年 | 3篇 |
1990年 | 5篇 |
1988年 | 6篇 |
1987年 | 6篇 |
1986年 | 3篇 |
1985年 | 2篇 |
1984年 | 8篇 |
1983年 | 2篇 |
1982年 | 3篇 |
1981年 | 2篇 |
1975年 | 1篇 |
1974年 | 3篇 |
1973年 | 1篇 |
1968年 | 2篇 |
1957年 | 1篇 |
1931年 | 1篇 |
1929年 | 1篇 |
1925年 | 1篇 |
1922年 | 1篇 |
排序方式: 共有584条查询结果,搜索用时 15 毫秒
1.
Leo S. Melchers Dave V. Thompson Ken B. Idler Saskia T. C. Neuteboom Ruud A. de Maagd Rob A. Schilperoort Paul J. J. Hooykaas 《Plant molecular biology》1988,11(2):227-237
The virulence loci play an essential role in tumor formation by Agrobacterium tumefaciens. Induction of vir gene expression by plant signal molecules is solely dependent on the virulence loci virA and virG. This study focused on the virA locus of the octopine type Ti plasmid pTi15955. The nucleic acid sequence of a 5.7-kilobase fragment encompassing virA was determined. Genetic analysis of this region revealed that virA contains one open reading frame coding for a protein of 91 639 daltons. Immunodetection with antibodies raised against a 35-kDa VirA fusion protein produced in E. coli identified the VirA product in wild-type Agrobacterium cells. Moreover, it is shown that the VirA protein is located in the cytoplasmic membrane fraction of Agrobacterium. These data confirm the proposed regulatory function of VirA whereby VirA acts as a membrane sensor protein to identify plant signal molecules in the environment. The proposed sensory function of VirA strikingly resembles the function of the chemotaxis receptor proteins of E. coli. 相似文献
2.
Jac M. M. J. G. Aarts Jan G. J. Hontelez Peter Fischer Ruud Verkerk Albert van Kammen Pim Zabel 《Plant molecular biology》1991,16(4):647-661
With a view to cloning the root-knot nematode resistance gene Mi in tomato by chromosome walking, we have developed a molecular probe for the tightly linked acid phosphatase-1 (Aps-1) locus. The acid phosphatase-1 allozyme (APS-11), encoded by the Aps-1
1 allele originating from Lycopersicon peruvianum, was purified to apparent homogeneity from tomato roots and suspension cells. Microsequencing of CNBr and tryptic peptides generated from APS-11 provided a partial amino acid sequence, which accounted for approximately 23% of the protein and revealed two stretches of homology with soybean proteins KSH3 and VSP27, comprising 22 matches within 26 amino acid residues. The partial amino acid sequence information enabled us to isolate a 2.4 kb genomic Aps-1
1 sequence by means of the polymerase chain reaction (PCR), primed by degenerate pools of oligodeoxyribonucleotides, synthesized on the basis of the amino acid sequences. Synthesis of the 2.4 kb PCR product was specific for genomic templates carrying the L. peruvianum Aps-1
1 allele. Crucial to the priming specificity and the synthesis of the 2.4 kb genomic sequence was the use of degenerate primer pools in which the number of different primer species was limited by incorporating deoxyinosine phosphate residues at three and four base ambiguities. In using cDNA as a template, a 490 bp sequence was obtained, indicating a high proportion of intron sequences in the 2.4 kb genomic Aps-1
1 sequence. The Aps-1
1 origin of the PCR product was confirmed by RFLP (restriction fragment length polymorphism) analysis, using both a chromosome 6 substitution line and a pair of nearly isogenic lines, differing for a small chromosomal region around the Aps-1/Mi loci. 相似文献
3.
Hans C. P. Matthijs Jan Maarten Van Steenbergen Ruud Kraayenhof 《Photosynthesis research》1985,7(1):59-67
The fluorescent probe 9-amino-6-chloro-2-methoxy acridine was used to study the energy transduction in the thylakoid and cell membranes of the cyanobacterium Plectonema boryanum. Apart from light-driven electron transfer, the dark endogenous respiration also leads to energization resulting in an ACMA fluorescence response, that is sensitive to the electron flow inhibitor 2, 5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, to the energy transfer inhibitors dicyclohexylcarbodiimide and venturicidine and to the uncoupler 5-chloro-3-t-butyl-2-chloro-4-nitrosalicylanilide.In spheroplasts, in which the cell membranes have lost their capacity to maintain a proton gradient, the respiration-and light-induced ACMA fluorescence changes (quenching) are similar to those in chloroplasts. In intact cells a combination of reversible quenching and enhancement of ACMA fluorescence was found. This dualistic behaviour is supposedly caused by an opposite orientation of the thylakoid and cell membranes. ACMA quenching at the level of the thylakoids was obtained either by respiratory or photosynthetic electron transfer and gave similar responses to those obtained in the spheroplasts. The slower ACMA fluorescence enhancement, only observed in cells with intact cell membranes, also evoked by both respiration and light-induced energization is sensitive to the compounds mentioned above and in addition to KCN.Our results support the view [8] that dark oxidation of substrates by O2 proceeds via the thylakoid membrane and terminates at a CN- sensitive oxidase located in the cell membrane which requires the involvement of a mobile cytoplasmic redox mediator.Abbreviations ACMA
9-amino-6-chloro-2-methoxy acridine
- chl a
chlorophyll a
- DBMIB
2, 5-dibromo-3-methyl-6-isopropyl-p-benzoquinone
- DCCD
dicyclohexylcarbodiimide
- DNP
dinitrophenol
- DNP-INT
dinitrophenyl ether of 2-iodo-4-nitrothymol
- FCCP
carbonylcyanide-p-trifluoro-methoxy phenylhydrazone
- S-13
5-chloro-3-t-butyl-2-chloro-4-nitrosalicylanilide
- tricine
N-2 (2-Hydroxy-1, 1-bis (hydroxymethyl) ethyl)-glycine
- Tris
Tris (hydroxymethyl) amino methane 相似文献
4.
5.
Associations between different agonistic and affiliative behavioural patterns of female domestic cats (Felis silvestris catus) were studied. In three groups of intact cats living in confinement frequencies of fourteen agonistic and affiliative behavioural
patterns were recorded. The technique of factor analysis (Principal Components Analysis followed by varimax rotation on a
dyads X behavioural patterns matrix) was used to detect clusters in these behavioural patterns. Five factors (or types of
interindividual relationships) were extracted per group. They accounted collectively for at least 77% of the total variance
present in the data. Although differences existed between groups with respect to behavioural patterns included in each factor,
four clusters of behaviours could be discriminated: (I) social rubbing, lordosis and rolling in front of partner (sexual behaviour),
(II) allogrooming, social sniffing, nosing, sniffing rear and treading (inspection-affiliative behaviour), (III) offensive
behaviour and staring, and (IV) defensive behaviour and staring. The role of these clusters in group living is discussed. 相似文献
6.
Jan Hendrickx Paul Coucke Marie-Claude Hors-Cayla G. Peter A. Smit Yoon S. Shin Johann Deutsch Jan Smeitink Ruud Berger Philip Lee John Fernandes Patrick J. Willems 《Genomics》1994,21(3)
We describe here a new type of X-linked liver glycogen storage disease. The main symptoms include liver enlargement and growth retardation. The clinical and biochemical abnormalities of this glycogenosis are similar to those of classical X-linked liver glycogenosis due to phosphorylase kinase deficiency (XLG). However, in contrast to patients with XLG, the patients described here have no reduced phosphorylase kinase activity in erythrocytes and leukocytes, and no enzyme deficiency could be found. Linkage analysis of four families with this X-linked type of liver glycogenosis assigned the disease gene to Xp22. Lod scores obtained with the markers DXS987, DXS207, and DXS999 were 3.97, 2.71, and 2.40, respectively, all at 0% recombination. Multipoint linkage analysis localized the disease gene between DXS143 and DXS989 with a maximum lod score of 4.70 at θ = 0, relative to DXS987. As both the classical XLG gene and the liver α-subunit of PHK (PHKA2) are also located in Xp22, this variant type of XLG may be allelic to classical XLG, and both diseases may be caused by mutations in PHKA2. Therefore, we propose to classify XLG as XLG type I (the classical type of XLG) and XLG type II (the variant type of XLG). 相似文献
7.
Johannes P. van Dijken Ruud A. Weusthuis Jack T. Pronk 《Antonie van Leeuwenhoek》1993,63(3-4):343-352
An overview is presented of the steady- and transient state kinetics of growth and formation of metabolic byproducts in yeasts.Saccharomyces cerevisiae is strongly inclined to perform alcoholic fermentation. Even under fully aerobic conditions, ethanol is produced by this yeast when sugars are present in excess. This so-called Crabtree effect probably results from a multiplicity of factors, including the mode of sugar transport and the regulation of enzyme activities involved in respiration and alcoholic fermentation. The Crabtree effect inS. cerevisiae is not caused by an intrinsic inability to adjust its respiratory activity to high glycolytic fluxes. Under certain cultivation conditions, for example during growth in the presence of weak organic acids, very high respiration rates can be achieved by this yeast.S. cerevisiae is an exceptional yeast since, in contrast to most other species that are able to perform alcoholic fermentation, it can grow under strictly anaerobic conditions.Non-Saccharomyces yeasts require a growth-limiting supply of oxygen (i.e. oxygen-limited growth conditions) to trigger alcoholic fermentation. However, complete absence of oxygen results in cessation of growth and therefore, ultimately, of alcoholic fermentation. Since it is very difficult to reproducibly achieve the right oxygen dosage in large-scale fermentations, non-Saccharomyces yeasts are therefore not suitable for large-scale alcoholic fermentation of sugar-containing waste streams. In these yeasts, alcoholic fermentation is also dependent on the type of sugar. For example, the facultatively fermentative yeastCandida utilis does not ferment maltose, not even under oxygen-limited growth conditions, although this disaccharide supports rapid oxidative growth. 相似文献
8.
Ecological compensation of the impacts of a road. Preliminary method for the A50 road link (Eindhoven-Oss, The Netherlands) 总被引:5,自引:0,他引:5
After years of efforts to avoid or reduce the impact of infrastructure projects on nature, the principle of ecological compensation has been incorporated in Dutch governmental policy. Ecological compensation aims to recover those ecological functions and natural values that still remain affected after maximum effort has been made to reduce the impact of the intervention (mitigation). The accepted aim of current policy is thus no-net-loss of area and quality by means of mitigative and compensatory measures. As part of the planning process for construction of a stretch of road in The Netherlands, viz., the A50 road link in the province of North Brabant, a Nature Compensation Plan (NCP) was required to be drawn up. This work has recently been completed by the Regional Directorate of Public Works and Water Management, the initiator of the intervention. The NCP, initially presented as a Draft Plan, was drawn up by the Regional Directorate using a preliminary method designed by the Centre of Environmental Science of Leiden University for deriving compensatory measures. After an opportunity for public comment, the Draft Plan was revised to form a Final NCP. This article describes, firstly, the preliminary method for deriving ecological compensatory measures. The method starts by quantifying the effects of habitat loss, habitat disturbance (by changes in noise emissions, in the water table and in outdoor recreational patterns), barrier action and fauna casualties. Following mitigation of impacts on nature, compensation for non-mitigable effects focuses successively on area size, derived from the impacts on breeding birds, and on area quality, derived from the habitat requirements of the vegetation and fauna groups affected by the road. Guidelines for identifying appropriate locations for compensation are also formulated. Secondly, the compensation method is applied to calculate the mitigative and compensatory measures for the A50 trajectory between Eindhoven and Oss. Thirdly, two comparisons are made: the Draft NCP is compared with the results of the preliminary method, and the Draft and the Final NCP are compared with one another in order to identify the role of the interest groups that played a major role in commenting on the Draft Plan. Finally, realization of the compensatory measures and development of the preliminary method itself are discussed. On the basis of the experience with the A50 case study, a more robust compensation method for road projects is to be developed. 相似文献
9.
Expression of insulin-like growth factor II (IGF-II) and histological changes in the thymus and spleen of transgenic mice overexpressing IGF-II 总被引:2,自引:0,他引:2
Leo T. M. Van der Ven Paul J. M. Roholl Maria G. Reijnen-Gresnigt Ruud J. Bloemen Sylvia C. van Buul-Offers 《Histochemistry and cell biology》1997,107(3):193-203
Previously, transgenic mice were constructed overexpressing human insulin-like growth factor II (IGF-II) under control of
the H2kb promoter. The IGF-II transgene was highly expressed in thymus and spleen, and these organs showed an increase in weight. In
the current study we have analyzed the sites of IGF-II mRNA expression, the distribution of IGF-II, IGF-I, and both IGF receptors,
and histomorphometrical changes in thymus and spleen. With in situ mRNA hybridization, expression of the IGF-II transgene
is found with high intensity in the thymic medulla and in the white pulp/marginal zone of the spleen, whereas there were scattered
positive cells in the thymic cortex and in the splenic red pulp. Hybridization was restricted to non-lymphocytic cells. Immunohistochemistry
revealed intense IGF-II peptide staining with the same distribution as IGF-II mRNA. There was additional intense IGF-II staining
of all elements in the splenic red pulp (including trabeculae) and diffuse, low level staining in the thymic cortex. These
findings were not observed in control mice. In the thymic medulla, most IGF-II producing cells co-labelled with keratin, whereas
a minor population also stained for the monocyte/macrophage marker MOMA-2. In the spleen, co-labelling of IGF-II producing
cells was found with MOMA-1 (marginal zone), or with the dendritic cell marker NLDC-145 (red pulp). IGF-I and both IGF receptors
were found in these organs in nearly all cell types, with a similar pattern in transgenic mice and in control animals. Histomorphometric
analysis revealed a marked increase of thymus cortex size and an increased trabecular size in the spleen. This suggests that
IGF-II overproduction induces local effects (auto/paracrine) in the thymic cortex, but not in the thymic medulla. Trabecular
growth in the spleen most likely is a distant effect (paracrine or endocrine) of IGF-II overproduction.
Accepted: 5 September 1996 相似文献
10.
Construction and characterization of the chimeric monoclonal antibody E48 for therapy of head and neck cancer 总被引:1,自引:0,他引:1
Ruud H. Brakenhoff Frank B. van Gog James E. Looney Marijke van Walsum Gordon B. Snow Guus A. M. S. van Dongen 《Cancer immunology, immunotherapy : CII》1995,40(3):191-200
Data from an ongoing clinical radioimmunoscintigraphy trial indicate that99mTc-labeled monoclonal antibody (mAb) E48 is highly capable of selectively targeting squamous cell carcinoma of the head and neck (HNSCC). The percentage of the injected dose per gram of tumor tissue was found to be high, rendering mAbE48 a promising candidate mAb for therapeutic purposes. We now describe the construction of a chimeric (moouse/human) mAb E48 by recombinant DNA technology. The genes encoding the variable domains of the heavy and light chain were cloned and ligated into experession vectors containing the human 1 heavy-chain gene and the human k lightchain gene respectively. Biological properties of the resulting chimeric mAb E48 were compared to the murine form in vitro and in vivo. The reactivities of chimeric (c)mAb and murine (m)mAb E48 with HNSCC, as assessed by immunohistochemical staining as well as immuno-blotting were shown to be similar. The affinity constant appeared to be 0.9×1010 M–1 and 1.6×1010 M–1 for the mmAb and cmAb respectively. The biodistribution of both antibodies was tested by simultaneous injection into nude mice bearing human HNSCC xenografts. cmAb E48 was found to be cleared more rapidly from the blood than mmAb E48, resulting in a 30% lower tumor uptake but similar tumor to non-tumor ratios, 3 days after injection. Moreover, it was shown that cmAb E48 is highly capable of lysing HNSCC targets in ADCC assays in vitro, whereas the mmAb appeared to be almost incative. These data indicate that cmAb E48 has potential as a targeting agent for the eradication of HNSCC in man. 相似文献