全文获取类型
收费全文 | 1148篇 |
免费 | 87篇 |
国内免费 | 81篇 |
出版年
2024年 | 1篇 |
2023年 | 11篇 |
2022年 | 9篇 |
2021年 | 14篇 |
2020年 | 30篇 |
2019年 | 38篇 |
2018年 | 25篇 |
2017年 | 30篇 |
2016年 | 32篇 |
2015年 | 28篇 |
2014年 | 51篇 |
2013年 | 83篇 |
2012年 | 43篇 |
2011年 | 96篇 |
2010年 | 44篇 |
2009年 | 99篇 |
2008年 | 60篇 |
2007年 | 56篇 |
2006年 | 59篇 |
2005年 | 74篇 |
2004年 | 52篇 |
2003年 | 63篇 |
2002年 | 46篇 |
2001年 | 45篇 |
2000年 | 34篇 |
1999年 | 29篇 |
1998年 | 17篇 |
1997年 | 11篇 |
1996年 | 14篇 |
1995年 | 24篇 |
1994年 | 27篇 |
1993年 | 24篇 |
1992年 | 14篇 |
1991年 | 21篇 |
1990年 | 3篇 |
1989年 | 3篇 |
1988年 | 2篇 |
1985年 | 1篇 |
1984年 | 2篇 |
1980年 | 1篇 |
排序方式: 共有1316条查询结果,搜索用时 31 毫秒
1.
《Critical reviews in biotechnology》2013,33(3):185-193
AbstractCultures of filamentous fungi that secrete significant amounts of exopolysaccharides are among the most difficult of fermentation fluids, presenting difficulties in the areas of aeration, agitation, mixing, and control that may in turn impact the physiology of the microorganism in an undesirable manner. The fungus Sclerotium glucanicum, which produces a potentially useful exopolysaccharide known as scleroglucan, illustrates many such difficulties. This review discusses in detail the range of physiological studies on the producing microorganism itself, including those concerning formation of “undesirable” byproducts, principally oxalate, but also, under certain conditions, other TCA cycle acids. In addition, the bioreactor technology in use for production of this type of biopolymer is discussed in relation to the difficulties such fluid types present. The potential of pneumatically agitated reactors for such production is evaluated, and the lack of fundamental studies on such reactors and on the hydrodynamics and mixing behavior of such complex fluids is pointed out. 相似文献
2.
Sung Ho Son Sung Mee Choi Kum Boo Choi Yun Hee Lee Dea Sook Lee Myung Suk Choi Young Goo Park 《Biotechnology and Bioprocess Engineering》1999,4(2):112-118
Calli were induced from 300,000 embryos isolated from immature to mature stage of seeds collected on late September from 14
elite trees. When the embryos were cultured onto plastic Petri-dish containing 20 mL of modified B5 basal medium supplemented
with 3% (w/v) sucrose, 500 mg/L casein hydrolysate, 250 mg/L myo-inositol, 0.5% (w/v) polyvinyl polypyrrolidon (PVPP), 2×MS
vitamins, 0.5 mg/L gibberellic acid, and 10 mg/L 2,4-D after 2 weeks of culture, yellowish-white calli were immediately formed
on the surfaces of embryos, and subcultured for 4 weeks in same culture medium. Because most of calli maintained for more
than 3 months were revealed differences in their colors, surface texture, and growth rate, visual selection was made for first
round screening. When the size of visually selected calli larger than 19 mm in their diameter were inoculated, persistent
proliferation was observed. Among the plating methods tested for the selection of rapid growing cell lines at single cell
and/or small cell aggregate level, 2-layer spread plating revealed as the best for single cell cloning. To enhance cell growth
and maintain high rate of viability for long-term culture of yew cells in bioreactor, final cell volume less than 50% in SCV
seemed to be the best. Time course study revealed that 30% of inoculum density was suitable for fed batch culture. Among the
tested conditional media, the rate of 1∶2 (old medium: fresh medium) was recorded at the best for cell growth. 相似文献
3.
Armin Bohmann Ralf Pörtner Jörg Schmieding Volker Kasche Herbert Märkl 《Cytotechnology》1992,9(1-3):51-57
A hybridoma cell was cultivated continuously in a membrane dialysis bioreactor with an integrated radial-flow fixed bed consisting of porous Siran® carriers over a period of 6 weeks. Antibodies accumulated to an average of 100 mg l?1, approx. 10 times more than in fixed bed cultures without dialysis membrane. Serum costs could be reduced about 85% due to an appropriate feeding strategy. Siran® carriers with 3–5 mm diameter showed an advantage compared to those with 1–2 mm diameter. For the 3–5 mm carrier the specific glucose uptake rate and the MAb production rate were constant, if the velocity was between 0.09 mm s?1 and 0.75 mm s?1. At higher velocities cells are washed out of the bed. Furthermore antibody consistency and cell stability were verified in long-term cultivations over a period of 96 days. From an estimation of the antibody concentration reachable with the reactor concept under optimal conditions a concentration 45 times higher compared to axial-flow fixed bed reactors and 11 times higher compared to stirred tank reactors can be expected. 相似文献
4.
In vitro multiplication in liquid culture of Syngonium contaminated with Bacillus spp. and Rathayibacter tritici 总被引:5,自引:0,他引:5
Contaminated Syngonium clusters were multiplied in an air lift bioreactor in liquid medium containing sucrose with the medium being circulated through a sterilizing filter. After 30 days, the culture in filtered medium produced 19.5 shoot initials per gram fresh weight of inoculum compared to 8.7 shoot initials produced in unfiltered medium. Transfer to an elongation medium with 30 mg l-1 Rifampicin produced shoots on 67% of the clusters, while transfer to elongation medium without Rifampicin poduced shoots on 40% of the clusters. Clusters grown for three subcultures in a reactor without medium filtration had lost their multiplication ability. Clusters grown for three subcultures in a reactor with filtration, however, continued to show a two-three fold increase in fresh weight and shoot production.Abbreviations MS
Murashige and Skoog 相似文献
5.
Abstract: Changes of the main hydrodynamic and oxygen transfer parameters during Aspergillus niger cultivation in an external-loop air-lift bioreactor of 200 dm3 operating capacity were investigated. The final average concentrations of biomass and citric acid obtained in batch fermentations were about 17 g 1-1 and 90 g 1-1 , respectively. Significant influence of the increasing biomass concentration on the rheological properties of the broth and operating parameters was found. Volumetric oxygen transfer coefficient. k L a , was found to be dependent on the apparent viscosity of the broth with an exponent of -0.984. 相似文献
6.
Plant regenerated organs such as shoots, bulbs, microtubers, corms, embryos, etc. have been successfully proliferated in the bioreactor. The use of a bioreactor leads to the development of technology suitable for large scale plant propagation. The basic construction and characteristics of various types of bioreactor systems are reviewed in relation to shoot and embryo cultures. A pilot scale 500 liter bioreactor system was applied to the production of large scale Stevia rebaudiana shoots.Abbreviations DW
dry weight
- EC
electrical conductivity
- FW
fresh weight
- ORP
oxidation-reduction potential 相似文献
7.
A stationary bubble-swarm has been used to aerate a mammalian cell culture bioreactor with an extremely low gas flow rate. Prolonging the residence time of the gas bubbles within the medium improved the efficiency of the gas transfer into the liquid phase and suppressed foam formation. An appropriate field of speed gradients prevented the bubbles from rising to the surface. This aeration method achieves an almost 90% transfer of oxygen supplied by the bubbles. Consequently, it is able to supply cells with oxygen even at high cell densities, while sparging with a gas flow of only 0.22·10–3–1.45·10–3 vvm (30–200 ml/h).The reactor design, the oxygen transfer rates and the high efficiency of the system are presented. Two repeated batch cultures of a rat-mouse hybridoma cell line are compared with a surface-aerated spinner culture. The used cell culture medium was serum-free, either with or without BSA and did not contain surfactants or other cell protecting agents. One batch is discussed in detail for oxygen supply, amino acid consumption and specific antibody production. 相似文献
8.
T. E. Tautorus M. M. Lulsdorf S. I. Kikcio D. I. Dunstan 《In vitro cellular & developmental biology. Plant》1994,30(1):58-63
Summary Embryogenic cell cultures ofPicea mariana (black spruce) and the species complexPicea glauca-engelmannii (interior spruce) were maintained either as suspensions in liquid medium in 250 or 500-ml-capacity shake-flasks, 7-liter-capacity
airlift or mechanically stirred bioreactors, or on agar-solidified medium. Cultures from each of the maintenance conditions
were subsequently transferred to agar-solidified LP medium containing 40μM (±) -abscisic acid for maturation into cotyledonary stage embryos. For both species, the highest maturation frequency resulted
from cultures grown in the airlift bioreactor. With black spruce cells grown in the airlift bioreactor containing LP medium
with 60 mM sucrose, a maximum of 7.1 g·liter−1 dry weight and 2892 embryos·ml−1 were obtained after 15 days. For interior spruce cells, a maximum dry weight of 5.9 g·liter−1 and 2698 embryos·ml−1 were obtained after 21 to 30 days. During culture over 2 wk, ammonia was almost completely utilized by both species, wherease
nitrate was depleted to 40% of the initial concentration. Sucrose was rapidly hydrolyzed to glucose and fructose by both species.
Black spruce cultures preferentially metabolized glucose, whereas interior spruce preferentially metabolized fructose. Improved
growth of interior spruce cells in mechanically stirred bioreactors occurred when cultured in LP medium with 60 mM fructose as the sole carbon source.
NRCC no. 36479 相似文献
9.
A two-stage culture strategy was studied for continuous high-level production of a foreign protein in the chemically inducible T7 expression system. The first stage is dedicated to the maintenance of plasmid-bearing cells and the second stage to the target protein synthesis by induction of cells coming from the first stage. On entering the second stage, recombinant cells undergo a gradual induction of the target gene expression. These plasmid-bearing cells experience dynamic changes in intracellular compositions and specific growth rates with their individual residence times. Therefore, the overall cultural characteristics in the production stage are really averages of the contributions from the various cells with different residence times. The behavior of the two-stage culture is described by a model, which accounts for dynamic variations of cell growth and protein synthesis rates with cell residence times. Model simulations were compared with experimental results at a variety of operating conditions such as inducer concentration and dilution rate. This model is useful for understanding the behavior of two-stage continuous cultures. (c) 1993 John Wiley & Sons, Inc. 相似文献
10.
This article demonstrates the successful in situ real-time monitoring of the cell concentration of Perilla frutescens in a bioreactor by using a laser turbidimeter. It was found that turbidity measurements at 780 nm with the laser sensor were hardly affected by the red color of the anthocyanin produced by P. frutescens cells, nor by the aeration rate or agitation speed within the ranges investigated. There was an excellent linear relationship, with a correlation coefficient (r(2)) higher than 0.99, between the sensor's response and the cell concentration. The whole growth stage of the cells, i.e., lag, logarithmic, and stationary phases, in bioreactor cultivations, could be satisfactorily estimated on-line by means of the in situ turbidimeter. However, during the declining phase of the cells, an apparent deviation was observed between the on-line estimations and off-line measurements of cell concentrations by dry cell weight, while the wet cell weight could be estimated by the same turbidimeter system. We found that this deviation was caused by a decrease in the cell density due to an increase of the individual cell volume and a decrease of the cell dry weight during the declining phase. (c) 1993 John Wiley & Sons, Inc. 相似文献