全文获取类型
收费全文 | 4711篇 |
免费 | 312篇 |
国内免费 | 303篇 |
专业分类
5326篇 |
出版年
2024年 | 7篇 |
2023年 | 69篇 |
2022年 | 160篇 |
2021年 | 240篇 |
2020年 | 153篇 |
2019年 | 184篇 |
2018年 | 186篇 |
2017年 | 140篇 |
2016年 | 201篇 |
2015年 | 261篇 |
2014年 | 338篇 |
2013年 | 370篇 |
2012年 | 411篇 |
2011年 | 347篇 |
2010年 | 228篇 |
2009年 | 211篇 |
2008年 | 232篇 |
2007年 | 166篇 |
2006年 | 161篇 |
2005年 | 159篇 |
2004年 | 155篇 |
2003年 | 146篇 |
2002年 | 104篇 |
2001年 | 118篇 |
2000年 | 83篇 |
1999年 | 95篇 |
1998年 | 46篇 |
1997年 | 33篇 |
1996年 | 36篇 |
1995年 | 34篇 |
1994年 | 30篇 |
1993年 | 18篇 |
1992年 | 29篇 |
1991年 | 23篇 |
1990年 | 31篇 |
1989年 | 8篇 |
1988年 | 17篇 |
1987年 | 12篇 |
1986年 | 9篇 |
1985年 | 8篇 |
1984年 | 10篇 |
1983年 | 8篇 |
1982年 | 7篇 |
1981年 | 5篇 |
1980年 | 6篇 |
1979年 | 3篇 |
1978年 | 3篇 |
1977年 | 3篇 |
1975年 | 6篇 |
1974年 | 3篇 |
排序方式: 共有5326条查询结果,搜索用时 0 毫秒
11.
Mutational analysis of the Saccharomyces cerevisiae ABD1 gene: cap methyltransferase activity is essential for cell growth.
下载免费PDF全文
![点击此处可从《Molecular and cellular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
RNA (guanine-7-)-methyltransferase is the enzyme responsible for methylating the 5' cap structure of eukaryotic mRNA. The Saccharomyces cerevisiae enzyme is a 436-amino-acid protein encoded by the essential ABD1 gene. In this study, deletion and point mutations in ABD1 were tested for the ability to support growth of an abd1 null strain. Elimination of 109 amino acids from the N terminus had no effect on cell viability, whereas a more extensive N-terminal deletion of 155 residues was lethal, as was a C-terminal deletion of 55 amino acids. Alanine substitution mutations were introduced at eight conserved residues within a 206-amino-acid region of similarity between ABD1 and the methyltransferase domain of the vaccinia virus capping enzyme. ABD1 alleles H253A (encoding a substitution of alanine for histidine at position 253), T282A, E287A, E361A, and Y362A were viable, whereas G174A, D178A, and Y254A were either lethal or severely defective for growth. Alanine-substituted and amino-truncated ABD1 proteins were expressed in bacteria, purified, and tested for cap methyltransferase activity in vitro. Mutations that were viable in yeast cells had either no effect or only a moderate effect on the specific methyltransferase activity of the mutated ABD1 protein, whereas mutations that were deleterious in vivo yielded proteins that were catalytically defective in vitro. These findings substantiate for the first time the long-held presumption that cap methylation is an essential function in eukaryotic cells. 相似文献
12.
Optimization of affinity and ion-exchange chromatographic processes for the purification of proteins
This study documents several alternative approaches for the optimization of the ion-exchange and affinity chromatographic purification of proteins. In these approaches, the chromatographic process has been treated as a four-stage (adsorption, washing, elution, and regeneration) operation. Central to these investigations has been the elaboration of practical iterative procedures based on the use of theoretical models describing each of these stages. Predictions derived from these models have then been evaluated in terms of experimental data obtained using batch adsorption measurements in finite bath configurations and frontal breakthrough measurements with packed beds of different dimensions, containing nonporous and porous adsorbents of different selectivities and capacities for proteins. Commencing with the kinetic and distribution parameters derived from batch equilibrium measurements, the effect of the initial concentration of the target protein, the solid-liquid volume ratio, the superficial velocity and the column dimensions on the pressure drop, production rate, concentration profile, column utilization, and yield have been determined with packed beds. The potential of these iterative approaches to simplify the determination of key mass transfer and interaction parameters required for scale-up and economic optimization of chromatographic purifications of proteins has been examined using ion exchange, immobilized metal ion affinity, and triazine dye pseudo-affinity adsorbents of different selectivity and adsorption capacities. (c) 1996 John Wiley & Sons, Inc. 相似文献
13.
14.
15.
观察了hFPIL6/2对6.5Gyγ线照射NIH小鼠第10天造血功能恢复的影响。结果表明:照射小鼠连续4d给予hFPIL6/2250μg·kg-1·d-1,其脾重、CFU-8、骨髓有核细胞数及CM-CFU分别比对照组增加59.0%、278.5%、57.9%和138.2%,统计学处理均有显著差异;对此四项指标的改善也明显优于25μg组。另外,250μg剂量组小鼠外周血象30d的动态观察结果表明,hFPIL6/2不但能明显提高红细胞和血红蛋白的最低值,而且能使血小板的恢复提前。提示hFPIL6/2在促进血小板生成和促进红系造血方面可能具有良好的应用前景。 相似文献
16.
以草鱼ZC7901细胞株为材料,观察鱼类细胞从间期染色质到中期染色体的包装过程。主要通过(1)分裂期与间期细胞融合,诱导染色体早熟凝集;(2)染色体“伸长”处理;(3)培养细胞的低渗处理;(4)染色质辅展等方法,制作染色体标本,进行扫描和透射电镜观察。观察表明,鱼类染色质的基本结构与哺乳类细胞相同,也是直径约10nm的核丝。染色体的色装有两种形式:一种是多级螺旋化形成直径约300nm的染色单体, 相似文献
17.
18.
棉花枯萎病菌多聚半乳糖醛酸内切酶在pH大于7时不稳定,故对它进行多种化学修饰而又不影响其活性,必须在pHd小于7的体系中进行。本文报道将PGAUase在还原剂存在下,与稀酸处理的Sepharose 4B交联,获得较高活力的固定化酶。固定化酶催化动力学表明,最适pH为4,4,最适温度为55℃,在pH1至8.0范围内稳定。和溶液酶比较,对热稳定性提高,但对碱稳定性下降。以多聚半乳糖醛酸为底物,Km为0.27mmol/L,Vmax为66.67nmol/L·min,均大于溶液酶(Km=0.07mmol/L,Vmax=28.00nmol/L·min)。在pH4.8,30℃,聚半乳糖醛酸在固相酶的柱中循环水解不同的时间降解产物经圆盘电泳和等电聚焦测定,得到不同大小的寡糖片段混合物,证明固相酶和溶液酶的作用方式相同,同时使以酶解法制备一定大小的有生物活性的寡糖分子成为可能。 相似文献
19.
Yuheng Chen Yongbin Wu Yuran Dong Yao Li Zhiwei Ge Oduro George Gang Feng Lingfeng Mao 《Diversity & distributions》2023,29(2):232-243
Aim
Understanding how species' traits and environmental contexts relate to extinction risk is a critical priority for ecology and conservation biology. This study aims to identify and explore factors related to extinction risk between herbaceous and woody angiosperms to facilitate more effective conservation and management strategies and understand the interactions between environmental threats and species' traits.Location
China.Taxon
Angiosperms.Methods
We obtained a large dataset including five traits, six extrinsic variables, and 796,118 occurrence records for 14,888 Chinese angiosperms. We assessed the phylogenetic signal and used phylogenetic generalized least squares regressions to explore relationships between extinction risk, plant traits, and extrinsic variables in woody and herbaceous angiosperms. We also used phylogenetic path analysis to evaluate causal relationships among traits, climate variables, and extinction risk of different growth forms.Results
The phylogenetic signal of extinction risk differed among woody and herbaceous species. Angiosperm extinction risk was mainly affected by growth form, altitude, mean annual temperature, normalized difference vegetation index, and precipitation change from 1901 to 2020. Woody species' extinction risk was strongly affected by height and precipitation, whereas extinction risk for herbaceous species was mainly affected by mean annual temperature rather than plant traits.Main conclusions
Woody species were more likely to have higher extinction risks than herbaceous species under climate change and extinction threat levels varied with both plant traits and extrinsic variables. The relationships we uncovered may help identify and protect threatened plant species and the ecosystems that rely on them. 相似文献20.
在赣榆县厉庄以大金鸡菊、紫穗槐和红柳三种经济植物进行了地埂利用型式试验,取得了比较明显的水土保持效果和改善农田小环境的生态效益。栽植后第三年植物覆盖率达90%或100%;地面蒸发量减少15.2~17.7%;截留降水量为0.5~0.7mm;大气相对湿度提高3.7~5.4%;土壤养分也有不同程度的增加。经济效益估测每亩地埂可收到300~600元,比种黄豆高5~9倍。效益最好的是红柳,大金鸡菊次之,紫穗槐较低。 相似文献