全文获取类型
收费全文 | 15596篇 |
免费 | 1344篇 |
国内免费 | 1908篇 |
专业分类
18848篇 |
出版年
2024年 | 53篇 |
2023年 | 281篇 |
2022年 | 591篇 |
2021年 | 906篇 |
2020年 | 701篇 |
2019年 | 815篇 |
2018年 | 757篇 |
2017年 | 553篇 |
2016年 | 725篇 |
2015年 | 1040篇 |
2014年 | 1251篇 |
2013年 | 1283篇 |
2012年 | 1562篇 |
2011年 | 1434篇 |
2010年 | 905篇 |
2009年 | 727篇 |
2008年 | 807篇 |
2007年 | 743篇 |
2006年 | 604篇 |
2005年 | 527篇 |
2004年 | 386篇 |
2003年 | 303篇 |
2002年 | 280篇 |
2001年 | 191篇 |
2000年 | 180篇 |
1999年 | 174篇 |
1998年 | 120篇 |
1997年 | 114篇 |
1996年 | 117篇 |
1995年 | 96篇 |
1994年 | 89篇 |
1993年 | 66篇 |
1992年 | 85篇 |
1991年 | 55篇 |
1990年 | 57篇 |
1989年 | 50篇 |
1988年 | 35篇 |
1987年 | 27篇 |
1986年 | 28篇 |
1985年 | 29篇 |
1984年 | 15篇 |
1983年 | 14篇 |
1982年 | 13篇 |
1981年 | 5篇 |
1980年 | 4篇 |
1979年 | 10篇 |
1978年 | 4篇 |
1976年 | 9篇 |
1975年 | 8篇 |
1971年 | 3篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
91.
92.
Aptamer selection for the detection of Escherichia coli K88 总被引:2,自引:0,他引:2
In this study, the first group of single-stranded DNA aptamers that are highly specific to enterotoxigenic Escherichia coli (ETEC) K88 was obtained from an enriched oligonucleotide pool by the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) procedure, during which the K88 fimbriae protein was used as the target and bovine serum albumin as counter targets. These aptamers were applied successfully in the detection of ETEC K88. They were then grouped under different families based on the similarity of their secondary structure and the homology of their primary sequence. Four sequences from different families were deliberately chosen for further characterization by fluorescence analysis. Having the advantage of high sensitivity, fluorescence photometry was selected as single-stranded DNA quantification method during the SELEX process. Aptamers with the highest specificity and affinity were analyzed to evaluate binding ability with E. coli. Since ETEC K88 is the only type of bacterium that expressed abundant K88 fimbriae, the selected aptamers against the K88 fimbriae protein were able to specifically identify ETEC K88 among other bacteria. This method of detecting ETEC K88 by aptamers can also be applied to bacteria other than ETEC K88. 相似文献
93.
To investigate the regulatory mechanisms of sex expression in cucumber, morphological observations and biochemical analyses were carried out on inappropriate stamen development of female flowers of cucumber. It was found that developmental arrest of the inappropriate stamen mainly occurs at the anther primordium. This arrest is closely correlated with DNA damage, as detected by TUNEL assay, and might result from anther-specific DNase activation. It was also found that the DNA damage does not lead to cell degeneration, although chromatin condensation is observed in the anther primordia.Abbreviations DAPI 4,6-diamidine-2-phenylindole dihydrochloride - MTT 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide - PCD programmed cell death - TUNEL TdT-mediated dUTP nick-end labeling 相似文献
94.
Hepatitis B virus (HBV) core protein (HBc) is a major component of viral nucleocapsid and a multifunctional protein involved
in viral maturation and release. It is unstable and present in cells at low level because of K96 lysine residue, which is
a ubiquitin acceptor site. Np95/ICBP90-like RING finger protein (NIRF) has auto-ubiquitination activity which is the hallmark
of a ubiquitin ligase. In the present study, ubiquitin ligase, NIRF, binds to HBc and leads to the proteasome-mediated degradation
of HBc in vivo. NIRF down-regulates HBc protein level, resulting in the decrease of the amount of HBV particles in supernatant
of HepG2.2.15 cells. However knockdown of NIRF significantly increases endogenous HBc protein level, leading to HBV release.
The results reveal that NIRF interacts with HBc and promotes the degradation of HBc in vivo. The pathway of NIRF-mediated
ubiquitin–proteasome affects the release of HBV particles by controlling the amounts of HBc. It indicates that NIRF may participate
in the maturation of HBV. 相似文献
95.
CC Cheng N Lu CL Peng CC Chang FD Mai LY Chen MH Liao WM Wang J Chang 《Proteomics》2012,12(15-16):2584-2597
The survivals of gastric cancer (GC) patients are associated with early diagnosis and effective treatments. Therefore, it is urgent for the discovery of early GC biomarkers and tumor-targeting therapeutics. The aim of this study was to uncover putative tissue biomarkers of GC using 2D DIGE and then apply one of these specific markers in GC treatment. We found three putative biomarkers of GC with significant differences in expression level compared to adjacent normal tissue, including glucose-regulated protein 78 (GRP78) and glutathione s-transferase pi (GSTpi) with increased expression level, and alpha-1 antitrypsin (A1AT) with reduced expression level. The overexpressed GRP78 was used as a targeted protein for guiding the drugs to tumor cells, leading to more effective treatment for GC xenografts. Our results demonstrated that the designated GRP78-binding peptide based on the sequence, WIFPWIQL, was selectively prone to recognize and bind to GC MKN45 cells in vitro, and also improve the delivery efficiency of polymeric micelles-encapsulated drugs into tumor cells and displayed better therapeutic outcome in experimental animals. This strategy of GRP78-mediated drug targeting system may bring chemotherapeutic drugs with more precise targeting to tumor cells, leading to minimize side effects on patients after chemotherapy. 相似文献
96.
97.
The biological effects of rare-earth ions on the organism have been studied using Pr3+ as a probe ion and Escherichia coli cell as a target. Atomic force microscopy (AFM) observation of the surface of E. coli cells shows that the presence of Pr3+ substantially changes the structure of the outer membrane. By induced coupled plasma-mass spectrometry (ICP-MS), more Cu2+ was found in the cells grown in the presence of Pr3+, indicating changes of cell permeability. Using energy dispersive X-ray spectroscopy (EDX), Ca2+ is found on the outer surface of the original cell. It is proposed that Pr3+ can replace Ca2+ from the binding sites because of their close ionic radii and similar ligand speciality. 相似文献
98.
Jun Feng Weixia Gao Yanyan Gu Wei Zhang Mingfeng Cao Cunjiang Song Peng Zhang Min Sun Chao Yang Shufang Wang 《Applied microbiology and biotechnology》2014,98(14):6397-6407
Poly-γ-glutamic acid (γ-PGA) is an important biopolymer with greatly potential in industrial and medical applications. In the present study, we constructed a metabolically engineered glutamate-independent Bacillus amyloliquefaciens LL3 strain with considerable γ-PGA production, which was carried out by single, double, and triple markerless deletions of three degradation genes pgdS, ggt, and cwlO. The highest γ-PGA production (7.12 g/L) was obtained from the pgdS and cwlO double-deletion strain NK-pc, which was 93 % higher than that of wild-type LL3 strain (3.69 g/L). The triple-gene-deletion strain NK-pgc showed a 28 % decrease in γ-PGA production, leading to a yield of 2.69 g/L. Furthermore, the cell morphologies of the mutant strains were also characterized. The cell length of cwlO deletion strains NK-c and NK-pc was shorter than that of the wild-type strain, while the ggt deletion strains NK-g, NK-pg, NK-gc, and NK-pgc showed longer cell lengths. This is the first report concerning the markerless deletion of γ-PGA degradation genes to improve γ-PGA production in a glutamate-independent strain and the first observation that γ-glutamyltranspeptidase (encoded by ggt) could be involved in the inhibition of cell elongation. 相似文献
99.
Conotoxins are a diverse array of small peptides mostly with multiple disulfide bridges. These peptides become an increasing significant source of neuro-pharmacological probes and drugs as a result of the high selectivity for ion channels and receptors. Usually, the analogue of natural conotoxins is produced by means of chemical synthesis. Here, we present a simple and fast strategy of producing disulfide-rich conotoxins via recombinant expression. By fused with thioredoxin and His tag, a novel O-superfamily conotoxin lt7a was successfully expressed in Escherichia coli and purified, resulting in a high yield of recombinant lt7a about 6 mg/l. The purity of target protein is up to 95% as identified by HPLC results. Whole cell patch-clamp recording revealed that the new conotoxin blocked voltage-sensitive sodium channels in rat dorsal root ganglion neurons, indicating it might be a novel microO-conotoxin. 相似文献
100.
番木瓜环斑病毒复制酶(亚基)基因的克隆、序列分析及其植物表达载体的构建 总被引:3,自引:0,他引:3
近二年报道,在TMV⑴、PVX⑵、PEBV⑶和CMV⑷等病毒中,利用病毒编码的复制酶(亚基)基因或相关cDNA片段转化烟草,工程植株获得绝对或极高的病毒抗性。大量研究认为:马铃薯Y病毒组的核内含体大分子量蛋白(Nib)是依赖于RNA的RNA复制酶(或核心亚基),Nlb与上述病毒的复制酶有广泛的同源保守区⑸。因此,Nib基因的克隆,不但在植物抗病基因工程方面,而且在马铃薯Y病毒组基因组的复制研究方面.均有重要的意义。本文以马铃薯Y病毒组的重要成员番木瓜环斑病毒的华南强株系(PRSV—sM)为材料,克隆了PRSV的Nib基因,并完成其全序列测定和植物表达载体的构建,为探索马铃薯Y病毒组复制酶可能介导的抗病性打下了基础。 相似文献