全文获取类型
收费全文 | 410篇 |
免费 | 16篇 |
国内免费 | 18篇 |
专业分类
化学 | 129篇 |
综合类 | 12篇 |
数学 | 1篇 |
物理学 | 7篇 |
综合类 | 295篇 |
出版年
2023年 | 2篇 |
2022年 | 4篇 |
2021年 | 15篇 |
2020年 | 4篇 |
2019年 | 6篇 |
2018年 | 4篇 |
2017年 | 10篇 |
2016年 | 9篇 |
2015年 | 14篇 |
2014年 | 11篇 |
2013年 | 12篇 |
2012年 | 20篇 |
2011年 | 18篇 |
2010年 | 13篇 |
2009年 | 16篇 |
2008年 | 18篇 |
2007年 | 25篇 |
2006年 | 33篇 |
2005年 | 28篇 |
2004年 | 29篇 |
2003年 | 20篇 |
2002年 | 24篇 |
2001年 | 18篇 |
2000年 | 13篇 |
1999年 | 17篇 |
1998年 | 13篇 |
1997年 | 7篇 |
1996年 | 9篇 |
1995年 | 8篇 |
1994年 | 8篇 |
1993年 | 4篇 |
1992年 | 4篇 |
1991年 | 3篇 |
1990年 | 1篇 |
1989年 | 2篇 |
1988年 | 1篇 |
1983年 | 1篇 |
排序方式: 共有444条查询结果,搜索用时 93 毫秒
11.
将活化癌基因T24-ras的全cDNA序列正向插入真核载体pMAMneo,构建成重组质粒pMAMneo-T24-ras.将该质粒转染NIH3T3细胞,通过药物筛选,建成细胞系3T3(T24).Southern杂交证明外源T24-ras基因已整合于受体细胞染色体中.3T3(T24)细胞表现出形态学方面的明显变化:具失去接触抑制能力,且在裸鼠体内致瘤等恶性行为.本文构建的T24-ras基因真核表达重组体和建立的转化细胞系可用于肿瘤的诊断、预防、治疗及抗肿瘤药物的筛选、评估等研究. 相似文献
12.
大肠杆菌植酸酶appA的融合表达及耐热性研究 总被引:2,自引:0,他引:2
提取大肠杆菌(SUGL)基因组DNA,通过PCR方法从基因组扩增得到植酸酶基因ap-pA,测序结果表明该基因的ORF读框包含1440个核苷酸.将该基因重组于大肠杆菌表达载体pET-32a(+)中,导入大肠杆菌BL21(DE3),构建工程菌BL21(DE3)-pET32a-appA.对表达条件进行了优化,在30℃下,以0.1mmol/L IPTG诱导表达植酸酶,表达量达到10%以上.在37℃,用钒钼酸铵法测定了表达的植酸酶活性为40740.7U/g,同时观察不同加热温度对植酸酶活性的影响程度.发现融合表达的 相似文献
13.
玉米矮花叶病在世界范围内广泛分布,且危害十分严重.因此正确认识玉米(Zea mays L.)对矮花叶病的抗性机制非常必要.基于此,本研究利用遗传差异较大的Mo17(高感玉米矮花叶病)和黄早四(高抗玉米矮花叶病)为亲本采用一粒传法构建了F9代重组自交系(Recombinant inbred line,RIL)分离群体,共256系.通过人工接甘蔗花叶病毒MDB株系(Sugarcane mosaic virus strain MDB, SCMV-MDB)对该分离群体进行了抗病性分析,统计了发病率和病情指数两项指标,根据这两项指标的频数分布图可知:玉米对矮花叶病的抗性主要由主基因控制,但不能排除还有微效基因的可能性. 相似文献
14.
类人胶原蛋白基因工程菌高密度发酵过程控制 总被引:1,自引:0,他引:1
目的优化基因工程菌Escherichia coli BL 21生产人源型胶原蛋白的高密度发酵工艺。方法建立一套以溶解氧浓度和比生长速率为中心的控制策略,采用溶氧反馈或pH反馈调节补料速度。结果 OD600可达150,胶原蛋白的表达量为9g/L。结论 20%-30%饱和度的溶解氧和0.1- 0.2 h-1的比生长速率有利于细胞生长和产物表达,其与降低乙酸等有害代谢副产物的积累密切相关。 相似文献
15.
16.
Ionizable amino acids in protein‐based hydrogels can confer pH‐responsive behavior. Because elastin‐like polypeptides (ELPs) have an established sequence and can crosslink to form hydrogels, they are an ideal system for creating pH‐sensitive materials. This study examines different parameters that might affect pH‐sensitive behavior and characterizes the mechanical and physical properties between pH 3 and 11 of three ELP‐based crosslinked hydrogels. The first finding is that varying the amount of crosslinker affects the overall stiffness and resilience of the hydrogels but does not strongly affect water content, swelling ratio, or pH sensitivity. Second, the choice of two popular tag sequences, which vary in histidine and aspartic acid content, does not have a strong effect on pH‐sensitive properties. Last, selectively blocking lysine and tyrosine residues through acetylation significantly decreases the pH‐sensitive zeta potential. Acetylated hydrogels also demonstrate different behavior at low pH values with reduced swelling, reduced water content, and higher stiffness. Overall, this work demonstrates that ELP hydrogels with ionizable groups are promising materials for environmentally‐responsive applications such as drug delivery, tissue engineering, and microfluidics. 相似文献
17.
Structural and Dynamic Characterization of the Molecular Hub Early Region 1A (E1A) from Human Adenovirus 下载免费PDF全文
Marcela Oliveira Nogueira Michele Salvi Dr. Talita Duarte Pagani Prof. Isabella C. Felli Prof. Roberta Pierattelli 《Chemistry (Weinheim an der Bergstrasse, Germany)》2016,22(37):13010-13013
The small‐DNA human adenovirus encodes one of the most versatile molecular hubs, the E1A protein. This protein is essential for productive viral infection in human cells and a vast amount of biologically relevant data are available on its interactions with host proteins. Up to now, however, no high‐resolution structural and dynamic information on E1A is available despite its important biological role. Among the different spliced variants of E1A, two are expressed at high level in the early stage of infection. These are 243 and 289 residues isoforms. Herein, we present their NMR characterization, showing that they are both highly disordered, but also demonstrate a certain heterogeneous behavior in terms of structural and dynamic properties. Furthermore, we present the characterization of the isolated domain of the longer variant, known as CR3. This study opens the way to understanding at the molecular level how E1A functions. 相似文献
18.
《高分子科学杂志,C辑:聚合物评论》2013,53(3):201-230
Super high modulus polyethylene fibers can be created by converting high molecular weight flexible PE chains into highly oriented and extended chain conformations. However, unlike polyethylene, aliphatic polyamides have very high cohesive energy and therefore cannot be easily drawn and highly oriented. This review addresses this fundamental problem by analyzing various novel approaches that can be used to suppress hydrogen bonding in these types of polyamides. Plasticization of such polymers with ammonia, iodine, salts, and Lewis acids, as well as dry spinning, wet spinning, and gel spinning, are discussed. Specialized techniques that involve vibrational zone drawing and annealing as well as laser heating zone drawing and annealing are also reviewed. Some of these methods definitely lead to remarkable improvements in initial modulus and other mechanical properties. The development of recombinant spider silk proteins as well progress in spinning these materials is also reported. The advantages and disadvantages of all of these processes are then summarized. 相似文献
19.
Narayan D. Chaurasiya Jacob Midiwo Pankaj Pandey Regina N. Bwire Robert J. Doerksen Ilias Muhammad Babu L. Tekwani 《Molecules (Basel, Switzerland)》2020,25(22)
A set of structurally related O-methylated flavonoid natural products isolated from Senecio roseiflorus (1), Polygonum senegalense (2 and 3), Bhaphia macrocalyx (4), Gardenia ternifolia (5), and Psiadia punctulata (6) plant species were characterized for their interaction with human monoamine oxidases (MAO-A and -B) in vitro. Compounds 1, 2, and 5 showed selective inhibition of MAO-A, while 4 and 6 showed selective inhibition of MAO-B. Compound 3 showed ~2-fold selectivity towards inhibition of MAO-A. Binding of compounds 1–3 and 5 with MAO-A, and compounds 3 and 6 with MAO-B was reversible and not time-independent. The analysis of enzyme-inhibition kinetics suggested a reversible-competitive mechanism for inhibition of MAO-A by 1 and 3, while a partially-reversible mixed-type inhibition by 5. Similarly, enzyme inhibition-kinetics analysis with compounds 3, 4, and 6, suggested a competitive reversible inhibition of MAO-B. The molecular docking study suggested that 1 selectively interacts with the active-site of human MAO-A near N5 of FAD. The calculated binding free energies of the O-methylated flavonoids (1 and 4–6) and chalcones (2 and 3) to MAO-A matched closely with the trend in the experimental IC50′s. Analysis of the binding free-energies suggested better interaction of 4 and 6 with MAO-B than with MAO-A. The natural O-methylated flavonoid (1) with highly potent inhibition (IC50 33 nM; Ki 37.9 nM) and >292 fold selectivity against human MAO-A (vs. MAO-B) provides a new drug lead for the treatment of neurological disorders. 相似文献
20.
应用重组维甲酸X受体(RXR)基因酵母筛选南方某污水处理厂进水、不同工艺出水的维甲酸干扰活性;结合大鼠肝均浆(S9)体外代谢方法,检测样品间接维甲酸干扰活性.结果表明:进水、出水样品均不能诱导RXR介导酶活性,但在高富集倍数下,样品与空白对照相比显著抑制9-顺维甲酸诱导酶活性,表现出维甲酸拮抗效应;在添加S9体外代谢活化后,所有样品均检出类维甲酸干扰活性,部分样品还检测出抗维甲酸活性,表明城市污水样品中存在大量具有类/抗维甲酸干扰活性的化合物.研究结果证实,现行处理工艺对类/抗维甲酸干扰物有一定的去除效果;重组基因酵母结合S9体外代谢活化体系可用于水环境样品类/抗维甲酸干扰效应的快速筛选. 相似文献