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191.
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Transgenic Arabidopsis plants can accumulate polyhydroxybutyrate to up to 4% of their fresh weight 总被引:7,自引:0,他引:7
Bohmert K Balbo I Kopka J Mittendorf V Nawrath C Poirier Y Tischendorf G Trethewey RN Willmitzer L 《Planta》2000,211(6):841-845
Transgenic Arabidopsis thaliana (L.) Heynh. plants expressing the three enzymes encoding the biosynthetic route to polyhydroxybutyrate (PHB) are described.
These plants accumulated more than 4% of their fresh weight (≈40% of their dry weight) in the form of PHB in leaf chloroplasts.
These very high producers were obtained and identified following a novel strategy consisting of a rapid GC-MS analysis of
a large number of transgenic Arabidopsis plants generated using a triple construct, thus allowing the parallel transfer of all three genes necessary for PHB synthesis
in a single transformation event. The level of PHB produced was 4-fold greater than previously published values, thus demonstrating
the large potential of plants to produce this renewable resource. However, the high levels of the polymer produced had severe
effects on both plant development and metabolism. Stunted growth and a loss of fertility were observed in the high-producing
lines. Analysis of the metabolite composition of these lines using a GC-MS method that we have newly developed showed that
the accumulation of high levels of PHB was not accompanied by an appreciable change in either the composition or the amount
of fatty acids. Substantial changes were, however, observed in the levels of various organic acids, amino acids, sugars and
sugar alcohols.
Received: 2 February 2000 / Accepted: 31 March 2000 相似文献
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Utility of Green Fluorescent Nucleic Acid Dyes and Aluminum Oxide Membrane Filters for Rapid Epifluorescence Enumeration of Soil and Sediment Bacteria 总被引:14,自引:7,他引:7 下载免费PDF全文
High background fluorescence and unspecific staining hampered the epifluorescence enumeration of bacteria in 45% of the tested soil and sediment samples with 4′,6-diamidino-2-phenylindole (DAPI) and polycarbonate membrane filters. These problems of the determination of total cell counts can be circumvented by using green fluorescent high-affinity nucleic acid dyes and aluminum oxide membrane filters. Due to the bright staining of cells, we recommend SYBR Green II as dye. 相似文献
195.
Delphine S. Dupuis Christiane Palmier Francis C. Colpaert Petrus J. Pauwels 《Journal of neurochemistry》1998,70(3):1258-1268
Abstract: G protein activation mediated by serotonin 5-HT1A and 5-HT1B/D receptors in guinea pig brain was investigated by using quantitative autoradiography of agonist-stimulated [35S]GTPγS binding to brain sections. [35S]GTPγS binding was stimulated by the mixed 5-HT1A/5-HT1B/D agonist L694247 in brain structures enriched in 5-HT1A binding sites, i.e., hippocampus (+140 ± 14%), dorsal raphe (+70 ± 8%), lateral septum (+52 ± 12%), cingulate (+36 ± 8%), and entorhinal cortex (+34 ± 5%). L694247 caused little or no stimulation of [35S]GTPγS binding in brain regions with high densities of 5-HT1B/D binding sites (e.g., substantia nigra, striatum, central gray, and dorsal subiculum). The [35S]GTPγS binding response was antagonized by WAY100635 (10 µM) and methiothepin (10 µM). In contrast, the 5-HT1B inverse agonist SB224289 (10 µM) did not affect the L694247-mediated [35S]GTPγS binding response, and the mixed 5-HT1B/D antagonist GR127935 (10 µM) yielded a partial blockade. The distribution pattern of the [35S]GTPγS binding response and the antagonist profile suggest the L694247-mediated response in guinea pig brain to be mediated by 5-HT1A receptors. In addition to L694247, 8-hydroxy-2-(di-n-propylamino)tetralin, and flesinoxan also stimulated [35S]GTPγS binding; their maximal responses varied between 46 and 52% compared with L694247, irrespective of the brain structure being considered. Sumatriptan, rizatriptan, and zolmitriptan (10 µM) stimulated [35S]GTPγS binding in the hippocampus by 20–50%. Naratriptan, CP122638, and dihydroergotamine stimulated [35S]GTPγS binding to a similar level as L694247 in hippocampus, lateral septum, and dorsal raphe. It appears that under the present experimental conditions, G protein activation through 5-HT1A but not 5-HT1B/D receptors can be measured in guinea pig brain sections. 相似文献
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Whole‐cell biotransformation of oleanolic acid by free and immobilized cells of Nocardia iowensis: Characterization of new metabolites 下载免费PDF全文
Benjamin Ludwig Doris Geib Christiane Haas Juliane Steingroewer Thomas Bley Kai Muffler Roland Ulber 《Engineering in Life Science》2015,15(1):108-115
In this study, Nocardia iowensis was used to transform oleanolic acid (OA) into oleanane derivatives. The first derivative, which was found after 24 h of cultivation, was the known and already described OA methyl ester. After 1 week, two other derivatives (oleanonic acid methyl ester and an unknown metabolite) were identified as new products of a biotransformation by N. iowensis. These oleanane metabolites were characterized by HPLC, HPLC‐ESI‐MS, and HPLC‐1H NMR spectroscopy. The biotransformation was performed by suspended and immobilized cells (ICs) of N. iowensis. Cells immobilized in alginate beads were used in order to prepare a continuous process. The substrate uptake of free and ICs was similar, whereas the peak area of OA methyl ester of the ICs was only about 10% of the native cells. However, the final product (oleanonic acid methyl ester) concentrations were similar in both approaches, whereas the unknown metabolite 3 was only detected transiently in the medium of ICs. Based on these results, a new biosynthetic pathway for the biotechnological production of oleanonic acid methyl ester is proposed. 相似文献
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