全文获取类型
收费全文 | 618篇 |
免费 | 27篇 |
专业分类
645篇 |
出版年
2023年 | 4篇 |
2021年 | 11篇 |
2018年 | 6篇 |
2017年 | 8篇 |
2016年 | 13篇 |
2015年 | 17篇 |
2014年 | 28篇 |
2013年 | 22篇 |
2012年 | 41篇 |
2011年 | 32篇 |
2010年 | 36篇 |
2009年 | 21篇 |
2008年 | 32篇 |
2007年 | 34篇 |
2006年 | 30篇 |
2005年 | 27篇 |
2004年 | 22篇 |
2003年 | 20篇 |
2002年 | 25篇 |
2001年 | 6篇 |
2000年 | 7篇 |
1999年 | 7篇 |
1998年 | 6篇 |
1997年 | 7篇 |
1996年 | 6篇 |
1995年 | 5篇 |
1994年 | 11篇 |
1993年 | 6篇 |
1992年 | 11篇 |
1990年 | 7篇 |
1989年 | 7篇 |
1987年 | 7篇 |
1986年 | 4篇 |
1984年 | 6篇 |
1981年 | 3篇 |
1980年 | 3篇 |
1978年 | 7篇 |
1977年 | 3篇 |
1976年 | 3篇 |
1975年 | 3篇 |
1974年 | 4篇 |
1965年 | 5篇 |
1964年 | 4篇 |
1962年 | 4篇 |
1958年 | 3篇 |
1957年 | 6篇 |
1956年 | 3篇 |
1955年 | 3篇 |
1951年 | 3篇 |
1949年 | 4篇 |
排序方式: 共有645条查询结果,搜索用时 0 毫秒
21.
Zhinxin Zhao Roberto Gambari Kenneth Ka-Ho Lee Stanton Hon-Lung Kok Raymond Siu-Ming Wong Fung-Yi Lau Johnny Cheuk-On Tang Kim-Hung Lam Chor-Hing Cheng Desmond Kwok Po Hau Chung-Hin Chui Wai-Yeung Wong Wai-Kwok Wong 《Bioorganic & medicinal chemistry letters》2013,23(8):2373-2376
We explore the possible cellular cytotoxic activity of an amphiphilic silicon(IV) phthalocyanine with axially ligated rhodamine B under ambient light experimental environment as well as its in vivo antitumour potential using Hep3B hepatoma cell model. After loading into the Hep3B hepatoma cells, induction of cellular cytotoxicity and cell cycle arrest were detected. Strong growth inhibition of tumour xenograft together with significant tumour necrosis and limited toxicological effects exerted on the nude mice could be identified. 相似文献
22.
23.
Promoter engineering to optimize recombinant periplasmic Fab′ fragment production in Escherichia coli 下载免费PDF全文
Desmond M. Schofield Alex Templar Joseph Newton Darren N. Nesbeth 《Biotechnology progress》2016,32(4):840-847
Fab’ fragments have become an established class of biotherapeutic over the last two decades. Likewise, developments in synthetic biology are providing ever more powerful techniques for designing bacterial genes, gene networks and entire genomes that can be used to improve industrial performance of cells used for production of biotherapeutics. We have previously observed significant leakage of an exogenous therapeutic Fab’ fragment into the growth medium during high cell density cultivation of an Escherichia coli production strain. In this study we sought to apply a promoter engineering strategy to address the issue of Fab’ fragment leakage and its consequent bioprocess challenges. We used site directed mutagenesis to convert the Ptac promoter, present in the plasmid, pTTOD‐A33 Fab’, to a Ptic promoter which has been shown by others to direct expression at a 35% reduced rate compared to Ptac. We characterized the resultant production trains in which either Ptic or Ptac promoters direct Fab’ fragment expression. The Ptic promoter strain showed a 25?30% reduction in Fab’ expression relative to the original Ptac strain. Reduced Fab’ leakage and increased viability over the course of a fed‐batch fermentation were also observed for the Ptic promoter strain. We conclude that cell design steps such as the Ptac to Ptic promoter conversion reported here, can yield significant process benefit and understanding with respect to periplasmic Fab’ fragment production. It remains an open question as to whether the influence of transgene expression on periplasmic retention is mediated by global metabolic burden effects or periplasm overcapacity. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:840–847, 2016 相似文献
24.
Detecting cell lysis using viscosity monitoring in E. coli fermentation to prevent product loss 下载免费PDF全文
Joseph M. Newton Desmond Schofield Joanna Vlahopoulou Yuhong Zhou 《Biotechnology progress》2016,32(4):1069-1076
Monitoring the physical or chemical properties of cell broths to infer cell status is often challenging due to the complex nature of the broth. Key factors indicative of cell status include cell density, cell viability, product leakage, and DNA release to the fermentation broth. The rapid and accurate prediction of cell status for hosts with intracellular protein products can minimise product loss due to leakage at the onset of cell lysis in fermentation. This article reports the rheological examination of an industrially relevant E. coli fermentation producing antibody fragments (Fab'). Viscosity monitoring showed an increase in viscosity during the exponential phase in relation to the cell density increase, a relatively flat profile in the stationary phase, followed by a rapid increase which correlated well with product loss, DNA release and loss of cell viability. This phenomenon was observed over several fermentations that a 25% increase in broth viscosity (using induction‐point viscosity as a reference) indicated 10% product loss. Our results suggest that viscosity can accurately detect cell lysis and product leakage in postinduction cell cultures, and can identify cell lysis earlier than several other common fermentation monitoring techniques. This work demonstrates the utility of rapidly monitoring the physical properties of fermentation broths, and that viscosity monitoring has the potential to be a tool for process development to determine the optimal harvest time and minimise product loss. © 2016 The Authors. Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers, 32:1069–1076, 2016 相似文献
25.
26.
Lee S. Webley Kyall R. Zenger Graham P. Hall Desmond W. Cooper 《European Journal of Wildlife Research》2007,53(1):40-46
European fallow deer are an introduced species classified as partly protected wildlife in Tasmania, Australia. Current management
practices are primarily governed under the Quality Deer Management regime, in which animals are harvested during designated
hunting seasons. Among populations, prominent morphological differences have been reported; however, the genetic relationship
of these populations has until now been poorly understood. Representative animals were sampled from three key areas across
their range and genotyped at ten polymorphic microsatellite loci to investigate genetic diversity, population structure, and
genetic bottlenecks. Allelic richness was low in all three populations and ranged between 2.20 and 2.49 alleles/locus. A genetic
bottleneck was detected in two of the three populations (P < 0.001). Population differentiation was evident between Lake Echo and Benham (q = 0.122; P < 0.001) and Benham and Connorville (q = 0.110; P < 0.001), but not between Lake Echo and Connorville (q = 0.0235), with individuals being identified as belonging to two genetic
clusters. The pattern of population differentiation from the three study populations suggests that deer from the western region
of their range are genetically distinct to those from the eastern region. This correlates with morphological variation within
Tasmanian fallow deer, in which differences between the regions maybe attributable to geographical barriers. 相似文献
27.
Matthew J. Desmond Liina Pajusalu Daniel W. Pritchard Tiffany A. Stephens Christopher D. Hepburn 《Journal of phycology》2019,55(4):936-947
Light availability is a fundamental factor that controls the productivity and distribution of macroalgae and is highly variable, both spatially and temporally, in subtidal coastal systems. Our comprehension of how macroalgae respond to such variability is a significant knowledge gap that limits our understanding of how light influences the structure and productivity of these environments. Here, we examined the pigment characteristics of individual species, and for the first time the whole community, within one low‐light, and one high‐light kelp‐forest system in southern New Zealand. The aim was to quantify the range of pigmentation seen within the two kelp‐forests which differed in irradiance regime. Light availability was 33% and 64% greater at the high‐light compared to the low‐light site at 2 and 10 m depth, respectively. Results suggested Phaeophyceae species at deeper depths in the low‐light site may be living at the edge of their photosynthetic ability and pigment synthesis appeared significantly restricted. Even with greater investment in the pigment fucoxanthin, biomass of Phaeophyceae species was significantly lower in the low‐light site. Highly pigmented Rhodophyceae species made a greater proportional contribution to community biomass within the low‐light site where they likely possessed a photosynthetic advantage. This work helps explain discrepancies in community structure between the two study sites and explores the complex relationship between irradiance and photoacclimation. The comparison of community pigment concentration holds potential as a tool for assessing the relative degree of photoacclimation occurring between sites and provides a proxy of photosynthetic cost under a specific light regime. 相似文献
28.
Peroxisomes are ubiquitous organelles that contain catalase (CAT) and an array of inducible enzymes that regulate aspects of lipid, purine, xenobiotic, eicosanoid, and phospholipid metabolism. Although peroxisomes are recognized as essential components in the cellular economy of microorganisms, plants, and mammals, little is known about their specialized functions in insect metabolism. Peroxisomal acyl-CoA oxidase (ACO) is a flavin-linked, H2O2-producing enzyme that regulates the β-oxidation of long chain fatty acids. We measured ACO and CAT activity in midgut tissues from worker honey bees, Apis mellifera, of known ages from free-flying colonies. The ACO activity peaked in young worker bees that digest and assimilate nutrients from pollen and from trophallaxis. As the bees aged, ACO activity declined. Conversely, CAT activity increased as the bees aged and reached its highest level in the oldest bees that were assayed. Isolated honey bee midguts were then fractionated using sucrose and Metrizamide (MET) density gradient centrifugation. Organelle-bound CAT activity equilibrated in sucrose at densities between 1.19 and 1.22, which are typical of spherical 1 μm peroxisomes. In the MET gradients, the organelle-bound CAT separated into two distinct fractions. The heavier fraction equilibrated at 1.21 and the lighter fraction at 1.15, a density commonly associated with microperoxisomes. These results support our ultrastructural and cytochemical data and suggest that the diverse functions of regionally specialized midgut epithelial cells lead to a heterogeneous population of peroxisomal organelles. ACO activity confirms the role of midgut peroxisomes in the intermediary metabolism of lipids and the increasing CAT activity suggests that the midgut epithelium may metabolize deleterious pro-oxidants of aerobic metabolism associated with foraging and senescence. © 1996 Wiley-Liss, Inc. 相似文献
29.
30.