全文获取类型
收费全文 | 493篇 |
免费 | 67篇 |
出版年
2022年 | 6篇 |
2021年 | 7篇 |
2018年 | 8篇 |
2017年 | 10篇 |
2016年 | 7篇 |
2015年 | 21篇 |
2014年 | 14篇 |
2013年 | 24篇 |
2012年 | 35篇 |
2011年 | 37篇 |
2010年 | 22篇 |
2009年 | 11篇 |
2008年 | 19篇 |
2007年 | 27篇 |
2006年 | 25篇 |
2005年 | 27篇 |
2004年 | 20篇 |
2003年 | 21篇 |
2002年 | 17篇 |
2001年 | 12篇 |
2000年 | 10篇 |
1999年 | 9篇 |
1998年 | 5篇 |
1997年 | 6篇 |
1996年 | 3篇 |
1995年 | 7篇 |
1994年 | 4篇 |
1993年 | 4篇 |
1992年 | 13篇 |
1991年 | 12篇 |
1990年 | 12篇 |
1989年 | 6篇 |
1988年 | 9篇 |
1986年 | 6篇 |
1985年 | 5篇 |
1984年 | 7篇 |
1983年 | 6篇 |
1982年 | 5篇 |
1981年 | 11篇 |
1980年 | 6篇 |
1977年 | 6篇 |
1974年 | 5篇 |
1973年 | 5篇 |
1971年 | 2篇 |
1970年 | 2篇 |
1969年 | 2篇 |
1968年 | 2篇 |
1967年 | 3篇 |
1966年 | 4篇 |
1965年 | 2篇 |
排序方式: 共有560条查询结果,搜索用时 31 毫秒
21.
Recombinant hirudin (r-hirudin), unlike the naturally occurring leech protein, lacks a sulfate ester on Tyr-63 which reduces its binding affinity to thrombin by 3-10-fold. We demonstrate that nitration or iodination of Tyr-63 restores hirudin-thrombin affinity to levels similar to or exceeding that of the natural inhibitor. In contrast, nitration of Tyr-3 reduces the affinity of hirudin for thrombin. These chemical modifications results in multiple reaction products that are readily separated by reverse-phase HPLC. The mechanism of the observed changes in thrombin affinity may involve a reduction in the pK of the hydroxyl group of tyrosine due to substitution of the electrophilic iodo or nitro group on the phenyl ring, resulting in an increased negative charge at neutral pH. For Tyr-63, this effect mimics the sulfatotyrosine of natural hirudin, leading to an increased thrombin affinity at the anion-binding exosite. For Tyr-3, the increased polarity may destabilize its interaction within the apolar-binding site of thrombin. Substitution of the highly conserved Tyr-3 residue with Phe or Trp not only enables specific and quantitative chemical modification at Tyr-63 but also independently increases hirudin-thrombin affinity. Kinetic analysis of thrombin inhibition showed that enhanced binding by r-hirudin(nitro-Tyr-63) is due to an increase in the association rate between hirudin and thrombin whereas the reduced binding of r-hirudin(nitro-Tyr-3) results from a large increase in the dissociation rate. These observations indicate that specific segments within both the amino- and carboxy-terminal regions of hirudin interact with thrombin. 相似文献
22.
Thermal Denaturation of Native Striatal Tyrosine Hydroxylase: Increased Thermolability of the Phosphorylated Form of the Enzyme 总被引:5,自引:5,他引:0
Mitchell A. Lazar Roger J. W. Truscott Joachim D. Raese Jack D. Barchas 《Journal of neurochemistry》1981,36(2):677-682
Tyrosine hydroxylase was purified from bovine corpus striatum. The native enzyme had a half-life of 15 +/- 3 min at 50 degrees C. Phosphorylation of tyrosine hydroxylase with protein kinase purified from both corpus striatum and heart activated the enzyme, but activity was rapidly lost with additional preincubation of the enzyme at 30 degrees C. Thermal denaturation studies indicated that phosphorylated tyrosine hydroxylase had a half-life of 5 +/- 2 min at 50 degrees C 相似文献
23.
Expression of cycloheximide resistance in carrot somatic hybrids and their segregants 总被引:3,自引:1,他引:2 下载免费PDF全文
Cycloheximide resistance (CHr) was shown to be a function expressed in differentiated plant tissues, but not in unorganized callus tissues. A variant, WCH105, expressing CHr in the callus, as well as in regenerated plantlets, was isolated from a cell line derived from a wild carrot plant. The plantlets regenerated from WCH105 are green, but do not produce normal, dissected leaves. Protoplasts of WCH105 were fused with that of a cycloheximidesensitive (CHs) cell line derived from an albino, domesticated carrot. Hybrid selection was based on (1) irreversible growth inhibition of WCH105 protoplasts by iodacetamide, and (2) restoration of green plants producing dissected leaves.——Analysis of the CHr trait as an unselected marker in the callus cells of the somatic hybrids indicated that it behaved as a recessive. The combined recessive and resistant phenotype of this trait allowed the recovery of CHr segregants from CHs hybrids at a frequency of 10-4, 1000 times higher than the spontaneous frequency of CHr. The recovery of CHr somatic segregants confirmed the recessiveness of the CHr trait. 相似文献
24.
25.
The production of granulocytes and macrophages from progenitor cells in the bone marrow is controlled, in part, by a family of humoral regulators, termed colony stimulating factors (CSF). We have examined genetic factors controlling this process using in vitro cloning techniques. The inbred mouse strain LP/J showed elevated colony formation (CFU-C) in response to one subtype of CSF (G,M-CSF) compared to other strains of mice examined including the strain C57BL/6J. This variation resulted in a shift to the left of the CFU-C dose-response curve for LP/J. No difference between LP/J and C57BL/6J was seen with another subtype of CSF (CSF-1). Maximal CFU-C response was similar in the two mouse strains with both types of CSF, and mixing experiments with both types of CSF gave the same maximal level of colony formation as the individual CSF. (C57BL/6J X LP/J)F1 progeny exhibited a CFU-C dose-response curve to CSF-2 that was intermediate between the parental types, indicating additive inheritance. Genetic analysis of backcross progeny suggested that the variation in CFU-C response is probably determined by a single primary gene, although the variability of the colony formation assay has complicated interpretation of genetic studies. These results suggest that CSF-1 and G,M-CSF act independently on a single bone marrow progenitor cell population. The properties of the genetic variation for G,M-CSF response are consistent with an alteration in cellular receptors for G,M-CSF. 相似文献
26.
Fuente David Lazar Dusan Oliver-Villanueva Jose Vicente Urchueguía Javier F. 《Photosynthesis research》2021,147(1):75-90
Photosynthesis Research - In this work, we reconstructed the absorption spectrum of different Synechocystis sp. PCC 6803 optical strains by summing the computed signature of all pigments present in... 相似文献
27.
Gideon V. Praveen Kumar Lazar Mathew 《Computer methods in biomechanics and biomedical engineering》2013,16(3):345-347
Vascular support structures are important devices for treating valve stenosis. A large population of patients is treated for valvular disease and the preferred mode of treatment is percutaneous valve replacement. Stent devices are proving to be an improved technology in minimally invasive cardiac surgery. This new technology provides highly effective results at minimal cost and with a short duration of hospitalisation. Stents as a supporting structure for tissue valves have evolved over the years into remarkably useful and effective devices. During this process, a number of specific designs have come and gone, and a few have remained. Many design changes were successful, and many were not. This article describes the merits and demerits of various stent designs and details the specific reasons why a particular novel design is expected to be the most suitable implant during and after percutaneous aortic valve replacement. 相似文献
28.
Varintip Srinon Sunsiree Muangman Nithima Imyaem Veerachat Muangsombut Natalie R. Lazar Adler Edouard E. Galyov Sunee Korbsrisate 《Journal of microbiology (Seoul, Korea)》2013,51(4):522-526
Burkholderia pseudomallei, the causative agent of melioidosis, is a Gram-negative saprophytic bacterium capable of surviving within phagocytic cells. To assess the role of BopC (a type III secreted effector protein) in the pathogenesis of B. pseudomallei, a B. pseudomallei bopC mutant was used to infect J774A.1 macrophage-like cells. The bopC mutant showed significantly reduced intracellular survival in infected macrophages compared to wild-type B. pseudomallei. In addition, the bopC mutant displayed delayed escape from endocytic vesicles compared with the wild-type strain. This indicates that BopC is important, and at least in part, needed for intracellular survival of B. pseudomallei. 相似文献
29.
Zbigniew Lazar Tristan Rossignol Jonathan Verbeke Anne-Marie Crutz-Le Coq Jean-Marc Nicaud Małgorzata Robak 《Journal of industrial microbiology & biotechnology》2013,40(11):1273-1283
Yarrowia lipolytica requires the expression of a heterologous invertase to grow on a sucrose-based substrate. This work reports the construction of an optimized invertase expression cassette composed of Saccharomyces cerevisiae Suc2p secretion signal sequence followed by the SUC2 sequence and under the control of the strong Y. lipolytica pTEF promoter. This new construction allows a fast and optimal cleavage of sucrose into glucose and fructose and allows cells to reach the maximum growth rate. Contrary to pre-existing constructions, the expression of SUC2 is not sensitive to medium composition in this context. The strain JMY2593, expressing this new cassette with an optimized secretion signal sequence and a strong promoter, produces 4,519 U/l of extracellular invertase in bioreactor experiments compared to 597 U/l in a strain expressing the former invertase construction. The expression of this cassette strongly improved production of invertase and is suitable for simultaneously high production level of citric acid from sucrose-based media. 相似文献
30.
Jiaying Xu Hongbo Shi Magaye Ruth Hongsheng Yu Lissy Lazar Baobo Zou Cui Yang Aiguo Wu Jinshun Zhao 《PloS one》2013,8(8)