全文获取类型
收费全文 | 3083篇 |
免费 | 384篇 |
国内免费 | 1174篇 |
出版年
2024年 | 28篇 |
2023年 | 72篇 |
2022年 | 167篇 |
2021年 | 155篇 |
2020年 | 159篇 |
2019年 | 194篇 |
2018年 | 141篇 |
2017年 | 133篇 |
2016年 | 123篇 |
2015年 | 197篇 |
2014年 | 275篇 |
2013年 | 222篇 |
2012年 | 270篇 |
2011年 | 302篇 |
2010年 | 232篇 |
2009年 | 206篇 |
2008年 | 257篇 |
2007年 | 216篇 |
2006年 | 210篇 |
2005年 | 179篇 |
2004年 | 145篇 |
2003年 | 120篇 |
2002年 | 124篇 |
2001年 | 116篇 |
2000年 | 117篇 |
1999年 | 60篇 |
1998年 | 44篇 |
1997年 | 31篇 |
1996年 | 12篇 |
1995年 | 20篇 |
1994年 | 20篇 |
1993年 | 13篇 |
1992年 | 14篇 |
1991年 | 17篇 |
1990年 | 10篇 |
1989年 | 5篇 |
1988年 | 3篇 |
1985年 | 4篇 |
1984年 | 3篇 |
1983年 | 3篇 |
1982年 | 5篇 |
1981年 | 2篇 |
1979年 | 2篇 |
1964年 | 2篇 |
1956年 | 2篇 |
1955年 | 1篇 |
1954年 | 1篇 |
1950年 | 1篇 |
1949年 | 1篇 |
1948年 | 1篇 |
排序方式: 共有4641条查询结果,搜索用时 15 毫秒
11.
分布于细胞内线粒体及细胞质中的天门冬氨酸氨基转移酶(AST·EC·2·6·1·1)是人血清中含有的两种同工酶,分别称为AST-m和AST-c同工酶.AST-c电泳迁移率介于血清α-球蛋白与β-球蛋白之间.AST-m电泳迁移率相似于γ-球蛋白,琼脂糖凝胶电泳固蓝B染色法对m-AST检出率较低.用NBT显色法则可得到较好效果. 相似文献
12.
石化来源的聚对苯二甲酸乙二酯(polyethylene terephthalate,PET)被广泛用于矿泉水瓶、食品包装和纺织品等领域,因其在自然界中不易分解,大量使用后的PET废弃物造成了严重的环境污染与资源浪费。使用生物酶法对PET废弃物进行解聚,并对解聚产物进行升级循环利用是进行塑料污染治理的重要方向之一,其中关键的是PET水解酶的解聚效率。对苯二甲酸双(羟乙基)酯(bis(hydroxyethyl)terephthalate,BHET)是PET生物酶解的中间产物,其累积是限制PET水解酶催化效率的一个重要因素,BHET水解酶和PET水解酶的联用能提升PET的整体水解效率。来源于嗜热氢化杆菌(Hydrogenobacter thermophilus)的双烯内酯酶(HtBHETase)对BHET有显著水解效果,将该酶在大肠杆菌(Escherichia coli)中进行重组表达并纯化后,对其酶学性质进行了研究。结果显示,HtBHETase对短碳链的酯类如对硝基苯酚乙酸酯催化活性较高,HtBHETase以BHET为底物时的最适反应pH值和最适反应温度分别为5.0和55℃;该酶有较好的热稳定性,经80℃的条件处理1 h仍能保持80%以上活性,显示出了良好的热稳定性,HtBHETase有在PET塑料生物解聚中使用的潜力,本研究为推动生物酶法降解PET提供了新的参考。 相似文献
13.
Cambrian lobopodians–ancestors of extant onychophorans? 总被引:7,自引:0,他引:7
This paper is a review of published information on fossil lobopodians, with addition of observations and ideas based on new material. It is also an analysis of the phytogeny of the group, and presents a new classification. A character shared by three or four families, yet not seen before, is a pair of enlarged sclerites covering the head. This forms an argument for re-orienting Hallucigenia once again. This genus no longer being enigmatic, a corner-stone in Stephen Jay Gould's evidence for extinct phyla is therefore gone. It is suggested that the lobopodians, phylum Lobopodia, are arranged in two classes, the extinct Xenusia for marine forms, and the Onychophora for terrestrial forms. The marine lobopodians are morphologically much more diverse than the extant onychophorans, a condition expressed in the classification. New taxa are: Hallucigenia fortis sp. nov., the families Luolishaniidae, Cardiodictyidae and Onychodictyidae, and the new orders Archonychophora, Scleronychophora and Paronychophora. 相似文献
14.
The size distribution of insertions and deletions in human and rodent pseudogenes suggests the logarithmic gap penalty for sequence alignment 总被引:20,自引:0,他引:20
The size distributions of deletions, insertions, and indels (i.e., insertions or deletions) were studied, using 78 human processed pseudogenes and other published data sets. The following results were obtained: (1) Deletions occur more frequently than do insertions in sequence evolution; none of the pseudogenes studied shows significantly more insertions than deletions. (2) Empirically, the size distributions of deletions, insertions, and indels can be described well by a power law, i.e., f
k
= Ck
–b
, where f
k
is the frequency of deletion, insertion, or indel with gap length k, b is the power parameter, and C is the normalization factor. (3) The estimates of b for deletions and insertions from the same data set are approximately equal to each other, indicating that the size distributions for deletions and insertions are approximately identical. (4) The variation in the estimates of b among various data sets is small, indicating that the effect of local structure exists but only plays a secondary role in the size distribution of deletions and insertions. (5) The linear gap penalty, which is most commonly used in sequence alignment, is not supported by our analysis; rather, the power law for the size distribution of indels suggests that an appropriate gap penalty is w
k
= a + b ln k, where a is the gap creation cost and blnk is the gap extension cost. (6) The higher frequency of deletion over insertion suggests that the gap creation cost of insertion (a
i
) should be larger than that of deletion (a
d
); that is, a
i
– a
d
= In R, where R is the frequency ratio of deletions to insertions.
Correspondence to: W.-H. Li 相似文献
15.
激活淋巴细胞cDNA单链库的构建 总被引:1,自引:0,他引:1
介绍一种激活淋巴细胞cDNA单链库构建方法,此库具有良好的模板活性. 用PCR法从此库中克隆了多种中国人源性的细胞因子. 相似文献
16.
Purification and characterization of 2,6-dichloro-p-hydroquinone chlorohydrolase from Flavobacterium sp. strain ATCC 39723. 总被引:1,自引:0,他引:1
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The biochemistry of pentachlorophenol (PCP) degradation by Flavobacterium sp. strain ATCC 39723 has been studied, and two enzymes responsible for the conversion of PCP to 2,6-dichloro-p-hydroquinone (2,6-DiCH) have previously been purified and characterized. In this study, enzymatic activities consuming 2,6-DiCH were identified from the cell extracts of strain ATCC 39723. The enzyme was purified to apparent homogeneity by a purification scheme consisting of seven steps. Gel filtration chromatography showed a native molecular weight of about 40,000, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single protein of 42,500 Da. The purified enzyme converted 2,6-DiCH to 6-chlorohydroxyquinol (6-chloro-1,2,4-trihydroxybenzene), which was easily oxidized by molecular oxygen and hard to detect. The end product, 6-chlorohydroxyquinol, was detected only in the presence of a reductase and NADH in the reaction mixture. The enzyme dechlorinated 2,6-DiCH but not 2,5-DiCH. The enzyme required Fe2+ for activity and was severely inhibited by metal chelating agents. The optimal conditions for activity were pH 7.0 and 40 degrees C. The Kcat for 2,6-DiCH was 35 microM, and the kcat was 0.011 s-1. 相似文献
17.
Purification and Properties of Component B of 2,4,5-Trichlorophenoxyacetate Oxygenase from Pseudomonas cepacia AC1100 总被引:3,自引:1,他引:2
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Pseudomonas cepacia AC1100 degrades 2,4,5-trichlorophenoxyacetate (2,4,5-T), an herbicide and chlorinated aromatic compound. Although some progress has been made in understanding 2,4,5-T degradation by AC1100 by molecular analysis, little is known about the biochemistry involved. Enzymatic activity converting 2,4,5-T to 2,4,5-trichlorophenol in the presence of NADH and O(inf2) was detected in cell extracts of AC1100. Phenyl agarose chromatography of the ammonium sulfate-fractionated cell extracts yielded no active single fractions, but the mixing of two fractions, named component A and component B, resulted in the recovery of enzyme activity. Component B was further purified to homogeneity by hydroxyapatite and DEAE chromatographies. Component B had a native molecular weight of 140,000, and it was composed of two 49-kDa (alpha)-subunits and two 24-kDa (beta)-subunits. Component B was red, and its spectrum in the visible region had maxima at 430 and 560 nm (shoulder), whereas upon reduction it had maxima at 420 (shoulder) and 530 nm. Each mole of (alpha)(beta) heterodimer contained 2.9 mol of iron and 2.1 mol of labile sulfide. These properties suggest strong similarities between component B and the terminal oxygenase components of the aromatic ring-hydroxylating dioxygenases. Component A was highly purified but not to homogeneity. The reconstituted 2,4,5-T oxygenase, consisting of components A and B, converted 2,4,5-T quantitatively into 2,4,5-trichlorophenol and glyoxylate with the coconsumption of NADH and O(inf2). 相似文献
18.
19.
20.
Purification and characterization of a tetrachloro-p-hydroquinone reductive dehalogenase from a Flavobacterium sp. 总被引:5,自引:0,他引:5
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Tetrachloro-p-hydroquinone (TeCH) is the first intermediate in pentachlorophenol (PCP) degradation by Flavobacterium sp. strain ATCC 39723. We previously purified a PCP hydroxylase that oxidized PCP to TeCH. Subsequently, we identified the reductive dehalogenation of TeCH to 2,3,6-trichloro-p-hydroquinone and then to 2,6-dichloro-p-hydroquinone in a cell extract with the reduced form of glutathione as the reducing agent under anaerobic conditions. Here we report the purification of a TeCH reductive dehalogenase that reductively dehalogenated TeCH to trichlorohydroquinone and then to dichlorohydroquinone. The enzyme was purified by protamine sulfate treatment, ammonium sulfate fractionation, and phenyl-agarose, anion-exchange, and gel filtration column chromatographies. As determined by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses, the protein has a molecular weight of about 30,000; nondenaturing polyacrylamide gel electrophoresis analysis suggests that the native enzyme exists as a dimer. The enzyme used glutathione but not NADPH, NADH, dithiothreitol, or ascorbic acid as the reducing agent. The optimal pH was close to neutral. 相似文献