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121.
The effect of ferredoxin(BED) overexpression on benzene dioxygenase activity in Pseudomonas putida ML2. 总被引:3,自引:3,他引:0 下载免费PDF全文
H M Tan C L Joannou C E Cooper C S Butler R Cammack J R Mason 《Journal of bacteriology》1994,176(9):2507-2512
The benzene dioxygenase from Pseudomonas putida ML2 is a multicomponent complex comprising a flavoprotein reductase, a ferredoxin, and a terminal iron-sulfur protein (ISP). The catalytic activity of the isolated complex shows a nonlinear relationship with protein concentration in cell extracts, with the limiting factor for activity in vitro being ferredoxin(BED). The relative levels of the three components were analyzed by using 125I-labelled antibodies, and the functional molar ratio of ISP(BED), ferredoxin(BED), and reductase(BED) was shown to be 1:0.9:0.8, respectively. The concentration of ferredoxin(BED) was confirmed by quantitative electron paramagnetic resonance spectroscopy of the 2Fe-2S centers in ferredoxin(BED) and ISP(BED) of whole cells. These results demonstrate that the ferredoxin(BED) component is a limiting factor in dioxygenase activity in vitro. To determine if it is a limiting factor in vivo, a plasmid (pJRM606) overproducing ferredoxin(BED) was introduced into P. putida ML2. The benzene dioxygenase activity of this strain, measured in cell extracts, was fivefold greater than in the wild type, and the activity was linear with protein concentration in cell extracts above 2 mg/ml. Western blotting (immunoblotting) and electron paramagnetic resonance spectroscopic analysis confirmed an elevated level of ferredoxin(BED) protein and active redox centers in the recombinant strain. However, in these cells, the increased level of ferredoxin(BED) had no effect on the overall rate of benzene oxidation by whole cells. Thus, we conclude that ferredoxin(BED) is not limiting at the high intracellular concentration (0.48 mM) found in cells. 相似文献
122.
123.
Summary An enzyme-immobilized microplate for determination of linamarin was prepared by covalently linking cassava leaf linamarase
to the microplate. For linamarin determination, cassava roots were homogenised in 0.1 Mo-phosphoric acid and the filtrate adjusted to pH 6 with NaOH prior to adding into the wells. The cyanide released was then
determined spectrophotometrically. One nmol linamarin can be detected. The microplate method is suitable for analysis of large
number of samples and is useful for screening purposes. 相似文献
124.
Effect of shock-loading of heavy metals on total organic carbon and phosphate removal in an anaerobic-aerobic activated sludge process 总被引:1,自引:0,他引:1
The effect of shock-loading of zinc, copper and cadmium ions on the removal of total organic carbon (TOC) and phosphate in an anaerobic-aerobic activated sludge process was investigated. TOC removal was not sensitive to shock-loading of Zn2+ and Cd2+ ions, and complete removal was achieved even at 20 mg Zn2+/l and 20 mg Cd2+/l. However, with over 1 mg Cu2+/1 TOC removal efficiency decreased. PO
inf4
sup3-
removal, in contrast, was extremely sensitive to these metal ions, with the threshold being 1 mg Zn2+/l and 1 mg Cd2+/l. Higher concentrations adversely affected PO
inf4
sup3-
removal. Copper again proved detrimental; no PO
inf4
sup3-
removal was achieved even at 1 mg Cu/l. These results highlight the sensitivity of the removal efficiencies of TOC and PO
inf4
sup3-
to shock loadings of these heavy metals.Y.P. Ting is with the Department of Chemical Engineering, National University of Singapore, Kent Ridge, 0511, Singapore; H. Imai and S. Kinoshita are with the Department of Chemical Process Engineering, Hokkaido University, Sapporo 060, Japan. 相似文献
125.
Tan Agnes; van den Broek Leon; Bolscher Jan; Vermass Dirk Jan; Pastoors Liesbeth; van Boeckel Constant; Ploegh Hidde 《Glycobiology》1994,4(2):141-149
N-Alkylation of the -glucosidase inhibitor 1-deoxynojirimycin(dNM) dramatically increases its inhibitory potency (Tan etal., J. Biol. Chem., 266, 1450414510, 1991). However,the possibility of extending the alkyl chain to N-decyl-dNMis limited by an increase of detergent-like (amphiphilic) propertiesof long-chain alkylated dNM derivatives. Substitution of methylenegroups in the N-decyl chain by oxygen reduced the amphiphilicityof N-decyl-dNM derivatives, while retaining their superior inhibitoryproperties. In intact HepG2 cells, the compound N-7-oxadecyl-dNMwas found to result in the most pronounced retention of glucoseresidues on N-linked glycans. Permeabilization of the plasmamembrane with the bacterial toxin Streptolysin O improves theinhibitory properties of the derivatives N-3,6,9-trioxadecyl-,N-7,10,13-trioxatetradecyl-, N-3-oxadecyl- and N-7-oxadecyl-dNM,but not those of dNM. These observations suggest differencesin the mode of entry of the oxygen-substituted dNM derivativesin comparison with dNM. We observed that the dNM derivativeN-3,6,9-trioxadecyl-dNM, devoid of inhibitory activity in intactcells, was inhibitory in Streptolysh O-permeabilized cells.Thus, the permeability barriers posed by plasma membrane andendoplasmic reticulum membrane are not equivalent. The use ofa permeabilized cell system thus allows the elaboration of inhibitoryprinciples for novel bioactive compounds where study of theisolated enzymes may not be possible, and where intact cellsare not a suitable target due to permeability barriers. -glucosidase inhibition N-linked glycosylation oxygen-substituted N-decyl-dNM derivatives permeabilized cells 相似文献
126.
水分胁迫对小麦光系统Ⅱ的影响 总被引:10,自引:0,他引:10
水分胁迫可降低小麦叶绿体的室温荧光产量和Mg2+ 在两个光系统间的调节能力、叶片的可变荧光产量、可变荧光猝灭速率以及荧光上升互补面积,表明光系统Ⅱ受到了伤害。光系统Ⅱ氧化侧的人工电子供体DPC能部分恢复受到抑制的叶绿体可变荧光,说明水分胁迫对光系统Ⅱ的损伤部位不仅位于氧化侧,也可能在反应中心上 相似文献
127.
质体醌重组的D1/D2/Cytb559复合物的荧光衰减动力学和光破坏作用 总被引:3,自引:0,他引:3
光系统Ⅱ反应中心D1/D2/Cytb559 在分离纯化过程中失去了电子受体QA 和QB,人工合成的质体醌可以与D1/D2/Cytb559 复合物发生重组。癸基质体醌(DPQ)与D1/D2/Cytb599 复合物的重组导致该复合物的荧光强度下降及发射光谱蓝移,同时两个与光化学活性相关的长寿命(24 ns和73 ns)荧光衰减组分占整个荧光的百分数下降,这些结果表明DPQ作为Pheo- 的电子受体,限制了P680+ ·Pheo- 的电荷重组。DPQ 的加入对D1/D2/Cytb559复合物中Chla 分子的光破坏敏感性影响不大,但β-胡萝卜素在加入DPQ 之后可以被光照破坏,这个过程可能与β-胡萝卜素的生理功能相关。 相似文献
128.
研究了大蒜(Allium sativum L.)中期染色体的超微结构和RNP物质。常规染色表明,大部分染色体内部有低电子密度区,有的染色体中低电子密度区域较大而似孔洞。银染结果也证明了有大小不等的孔洞存在。Bernhard 染色显示,在染色体周边和染色体内部都有RNP分布。用NaOH 处理证明了Bernhard 染色法所显示的深染区确实含有RNA。RNP量的多少与EDTA 的分化时间呈负相关 相似文献
129.
130.
OBJECTIVE: To determine the rates and outcomes of diabetic end-stage renal disease (ESRD) among registered native people and non-native people in Saskatchewan. DESIGN: Retrospective population-based study using data from the Canadian Organ Replacement Registry. SETTING: Saskatchewan. PATIENTS: All patients with diabetic ESRD diagnosed between Jan. 1, 1981, and Dec. 31, 1990. MAIN OUTCOME MEASURES: Incidence rates of diabetic ESRD in the general population, rates of diabetic ESRD among patients with diabetes mellitus, nature of initial dialysis treatment, length of survival from start of dialysis, cause of death and renal transplant rates. RESULTS: The 10-year incidence rates of diabetic ESRD were higher among all age groups among registered native people than among non-native people. The overall relative risk ratio for native people was 16.2. When a higher prevalence of diabetes among native people was taken into account, native diabetic people were still seven times as likely as non-native diabetic people to manifest diabetic ESRD. The median survival from start of dialysis was under 2 years in both groups, but more native people died of stroke and more non-native people died of heart disease. Non-native diabetic people were more likely than native diabetic people to receive renal transplants. CONCLUSIONS: Although the overall incidence of diabetic ESRD in Saskatchewan is increasing, registered native people have a disproportionate risk for this serious complication. 相似文献