首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   23527篇
  免费   2788篇
  国内免费   8670篇
  2024年   89篇
  2023年   579篇
  2022年   986篇
  2021年   1314篇
  2020年   1143篇
  2019年   1418篇
  2018年   1077篇
  2017年   1026篇
  2016年   1137篇
  2015年   1547篇
  2014年   1925篇
  2013年   1875篇
  2012年   2505篇
  2011年   2251篇
  2010年   1656篇
  2009年   1692篇
  2008年   1871篇
  2007年   1757篇
  2006年   1622篇
  2005年   1357篇
  2004年   1063篇
  2003年   889篇
  2002年   817篇
  2001年   691篇
  2000年   599篇
  1999年   415篇
  1998年   235篇
  1997年   184篇
  1996年   142篇
  1995年   102篇
  1994年   101篇
  1993年   77篇
  1992年   93篇
  1991年   75篇
  1990年   80篇
  1989年   61篇
  1988年   49篇
  1987年   45篇
  1986年   63篇
  1985年   44篇
  1984年   38篇
  1983年   34篇
  1982年   41篇
  1981年   28篇
  1980年   13篇
  1979年   24篇
  1978年   16篇
  1977年   14篇
  1975年   13篇
  1959年   11篇
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
91.
小麦条锈菌毒性小种及其无毒性突变型侵染初期,是不亲和反应的小麦叶片内可翻译mRNA水平迅速增加,而呈亲和反应叶片的增加幅度小且滞后。同时前者的Poly(A+)-RNA水平高于未接种对照,后者低于对照。32P标记实验证实不亲和反应叶片Poly(A+)-RNA的合成增加早于亲和反应叶片。Poly(A+)-RNA体外翻译产物经SDS-PAGE分离后,放射自显影图谱显示一些多肽条带的35S-Met相对掺入量有定量差异。  相似文献   
92.
Studies of cell pellets: I. Electrical properties and porosity.   总被引:2,自引:1,他引:1  
I G Abidor  L H Li    S W Hui 《Biophysical journal》1994,67(1):418-426
Cell pellets formed by centrifugation provided a good system to study the osmotic behavior, electroporation, and interaction between cells. Rabbit erythrocyte pellets were used in this study because they were simpler than nucleated cells to model analytically. Structurally, cell pellets possessed properties of porous solid bodies and gels. Electrically, cell pellets were shown to behave as a parallel set of resistance, Rp, and capacitance, Cp. Information on pellet structures was obtained from electric measurements. The pellet resistance reflected the intercellular conductivity (porosity and gap conductivity), whereas the pellet capacitance depended mostly on membrane capacitance. The pellet resistance was more sensitive to experimental conditions. The intercellular gap distance can be derived from pellet porosity measurements, providing the cell volume and surface area were known. Rp increased and relaxed exponentially with time when centrifugation started and stopped; the cycles were reversible. When supernatants were exchanged with solutions containing hypotonic electrolytes or macromolecules (such as PEG) after the pellets were formed, complicated responses to different colloidal osmotic effects were observed. A transient decrease followed by a large increase of Rp was observed after the application of a porating electric pulse, as expected from a momentary membrane breakdown, followed by a limited colloidal-osmotic swelling of pelleted cells. The equilibrium values of Rp, Cp, pellet porosity, and intercellular distances were measured and calculated as functions of cell number, centrifugation force, and ionic strength of the exchanged supernatant. Thus, the structure and properties of cell pellets can be completely characterized by electrical measurements.  相似文献   
93.
Chinese Hamster Ovary (CHO) cells were fused by subjecting cell suspensions to an exponentially decaying electric pulse in the presence of polyethylene glycol (PEG), Dextran or Ficoll. PEG (MW 1,000, 3,350, 8,000, 10,000 and 18,500), Dextran (MW 71,200) and Ficoll (MW 400,000) were added to the pulsing medium. A single exponential electric pulse with peak field strength of 4 kV/cm, and a half-time of 0.72 msec was used. The combination of two techniques, PEG-induced fusion and electrofusion, resulted in highly efficient fusion of CHO cells. Fusion yields (FY) at different concentrations of these polymers were measured using phase-contrast microscopy. FY was highly dependent on the concentration of PEG in media, while the presence of Dextran and Ficoll had no influence on fusion yield. PEG with MW 8,000 was found to be the most effective in causing cell aggregation, and to give the highest FY (40%). An optimal concentration for fusion was found for PEG of each molecular weight. Diluting cells suspended in higher concentrations of PEG to these optimal concentrations after the pulse application regained the optimal FY. It was concluded that PEG-induced prepulse aggregation and moderate cell swelling immediately after the pulse were important factors in achieving high fusion yields.This work is supported by a grant GM-30969 from the National Institutes of Health. Traveling fellowship to N.G.S. was supported from Foundation Cyrill and Methodius and grant N-189 from MCES of Bulgaria.  相似文献   
94.
Summary Heavy metals in electroplating effluent inhibited specific methanogenic activity (SMA) of anaerobic starch-degrading granules. The SMA of granules on the degradation of starch were reduced by 50% when each gram of biomass was in contact individually with 105 mg of zinc, 120 mg of nickel, 180 mg of copper, 310 mg of chromium, or >400 mg of cadmium. Granules had higher toxicity-resistance than flocculent sludge, due to their layered structure.  相似文献   
95.
旋毛虫肌幼虫ES抗原的基因克隆及高效表达   总被引:7,自引:0,他引:7  
作者对编码旋毛虫肌幼虫ES抗原的部分结构基因进行了克隆、鉴定和表达。用RNA PCR技术直接从旋毛虫肌幼虫总RNA中反转录并扩增出0.7kh的靶DNA,酶切分析后将其克隆到融合表达载体pEx3lC中。SDS—PAGE电泳表明,含重组子的大肠杆菌能够表达出一分子量为37kDa的融合蛋白(P37),后者占菌体总蛋白的22%以上,并以包含体形式存在于菌体中。经对纯化后表达蛋白的ELlSA检测,证明它能被猪旋毛虫病阳性血清和抗旋毛虫单克隆抗体识别。研究结果揭示,重组蛋白P37对于研制旋毛虫病诊断抗原和免疫抗原具有潜在的应用价值。  相似文献   
96.
棉铃虫对氰戊菊酯抗性和敏感品系的选育   总被引:40,自引:9,他引:31  
吴益东  沈晋良 《昆虫学报》1994,37(2):129-136
用氰戊菊酯对来自阳谷的棉铃虫(YG)Heliothis armigera(Hubncr) 进行抗性晶系的筛选。在15代期间经过9代的室内选育,获得抗性品系(Fcn-R),抗性倍数高达2“3倍,筛选后F15代LD50值(24.1412μg/头)比筛选前F1代lD50值(0.2020μg/头)提高了119.5倍。对来自偃师的棉铃虫(YS)进行了连续两代单对筛选,得到敏感品系(Fen-S),敏感晶系的LD50值为0.0116μg/头,接近1983年东台敏感晶系的LD50值(0.0098μg/头)Fcn-R抗性晶系筛选前后分别测定了七种杀虫剂的剂量-死亡回归线,发现Fen-R抗性品系对溴氰菊酯[LD50(Fen-R)/LD50(YG)=5.2X] 和氯氰菊酯(2.5X)具有一定程度的交互抗性;而对功夫菊酯(0.66X),氯菊酯(0.89x)、灭多威(0.74X)及久效磷(1.5x)没有交互抗性。氰戊菊酯加Pb的增效试验结果表明棉铃虫对氰戊菊酯的抗性主要是由于多功能氧化酶的代谢作用。毒理学资料还暗示抗性为多因子(基因)的。  相似文献   
97.
对沙打旺的下胚轴、子叶、幼叶组织培养中脱分化细胞进行了超微结构观察,并着重讨论了细胞核的动态变化。脱分化细胞的细胞质中线粒体墙加,嵴明显;多聚核糖体增多;高尔基体增加;质体中积累淀粉。核仁与核内异染色质之间有一个动态过程。此过程暂称“核仁物质喷射“现象。在致有以下:1.核体出现,半嵌在增大的核仁上,核内异染色质沿核膜凝聚;2.异染色质移向核仁,并与核仁接触,核体消失,部分异质进入核仁;3.核仁物质  相似文献   
98.
鹅掌楸属花粉的超微结构研究及系统学意义   总被引:23,自引:4,他引:19  
  相似文献   
99.
枸杞果实Fe-SOD粗提液经硫酸铵盐析、离子交换柱层析及凝胶过滤,纯化到电泳单班点均一程度。纯化的Fe-SOD分子量为44.6kD,亚基分子量为22.0kD。金属元素分析表明,每分子酸含1个Fe原子。该酶在紫外区最大吸收值为278um。H2O2明显抑制该酶活性,KCN对酶活性无影响。该酶氨基酸组成与高等植物和蓝绿藻的Fe-SOD相似,但它具有较高甘氨酸,酸性氨基酸与碱性氨基酸比值高于高等植物而与低等植物及原核生物相近。  相似文献   
100.
鸡马立克氏病病毒B抗原片段在大肠杆菌中表达   总被引:1,自引:0,他引:1  
鸡马立克氏病病毒BamHI基因文库I3质粒中含有编码B抗原膜外Domain的DNA序列。经ScaI和SphI双酶酶解I3质粒,分离获得764bpDNA片段,并克隆进M13mp19中。DNA序列测定分析表明克隆片段为MDV-B抗原基因的494-1258bp部分序列。进一步分离NcoI-HindIII部分基因片(530bp),克隆于PLPromoter控制下的含有修饰型cIts857基因的表达载体中,  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号