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排序方式: 共有194条查询结果,搜索用时 15 毫秒
91.
Martin Mück-H?usl Manish Solanki Wenli Zhang Zsolt Ruzsics Anja Ehrhardt 《Nucleic acids research》2015,43(8):e50
Recombinant adenoviruses containing a double-stranded DNA genome of 26–45 kb were broadly explored in basic virology, for vaccination purposes, for treatment of tumors based on oncolytic virotherapy, or simply as a tool for efficient gene transfer. However, the majority of recombinant adenoviral vectors (AdVs) is based on a small fraction of adenovirus types and their genetic modification. Recombineering techniques provide powerful tools for arbitrary engineering of recombinant DNA. Here, we adopted a seamless recombineering technology for high-throughput and arbitrary genetic engineering of recombinant adenoviral DNA molecules. Our cloning platform which also includes a novel recombination pipeline is based on bacterial artificial chromosomes (BACs). It enables generation of novel recombinant adenoviruses from different sources and switching between commonly used early generation AdVs and the last generation high-capacity AdVs lacking all viral coding sequences making them attractive candidates for clinical use. In combination with a novel recombination pipeline allowing cloning of AdVs containing large and complex transgenes and the possibility to generate arbitrary chimeric capsid-modified adenoviruses, these techniques allow generation of tailored AdVs with distinct features. Our technologies will pave the way toward broader applications of AdVs in molecular medicine including gene therapy and vaccination studies. 相似文献
92.
Solanki PR Prabhakar N Pandey MK Malhotra BD 《Journal of molecular recognition : JMR》2008,21(4):217-223
Nucleic acid sensor based on polyaniline (PANI) has been fabricated by covalently immobilizing double stranded calf thymus (dsCT) DNA onto perchlorate (ClO(-) (4))-doped PANI film deposited onto indium-tin-oxide (ITO) glass plate using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) chemistry. These dsCT-DNA-PANI-ClO(4)/ITO and PANI-ClO(4)/ITO electrodes have been characterized using square wave voltammetry, electrochemical impedance, scanning electron microscopy (SEM) and Fourier-transform-infrared (FTIR) measurements. This disposable dsCT-DNA-PANI-ClO(4)/ITO bioelectrode, stable for about 4 months, can be used to detect cypermethrin (0.005 ppm) and trichlorfon (0.01 ppm) in 30 and 60 s, respectively. 相似文献
93.
Electrical detection of the temperature induced melting transition of a DNA hairpin covalently attached to gold interdigitated microelectrodes 下载免费PDF全文
Brewood GP Rangineni Y Fish DJ Bhandiwad AS Evans DR Solanki R Benight AS 《Nucleic acids research》2008,36(15):e98
The temperature induced melting transition of a self-complementary DNA strand covalently attached at the 5′ end to the surface of a gold interdigitated microelectrode (GIME) was monitored in a novel, label-free, manner. The structural state of the hairpin was assessed by measuring four different electronic properties of the GIME (capacitance, impedance, dissipation factor and phase angle) as a function of temperature from 25°C to 80°C. Consistent changes in all four electronic properties of the GIME were observed over this temperature range, and attributed to the transition of the attached single-stranded DNA (ssDNA) from an intramolecular, folded hairpin structure to a melted ssDNA. The melting curve of the self-complementary single strand was also measured in solution using differential scanning calorimetry (DSC) and UV absorbance spectroscopy. Temperature dependent electronic measurements on the surface and absorbance versus temperature values measured in solution experiments were analyzed assuming a two-state process. The model analysis provided estimates of the thermodynamic transition parameters of the hairpin on the surface. Two-state analyses of optical melting data and DSC measurements provided evaluations of the thermodynamic transition parameters of the hairpin in solution. Comparison of surface and solution measurements provided quantitative evaluation of the effect of the surface on the thermodynamics of the melting transition of the DNA hairpin. 相似文献
94.
Bioactive compounds from marine actinomycetes 总被引:1,自引:0,他引:1
Actinomycetes are one of the most efficient groups of secondary metabolite producers and are very important from an industrial
point of view. Among its various genera, Streptomyces, Saccharopolyspora, Amycolatopsis, Micromonospora and Actinoplanes are the major producers of commercially important biomolecules. Several species have been isolated and screened from the
soil in the past decades. Consequently the chance of isolating a novel actinomycete strain from a terrestrial habitat, which
would produce new biologically active metabolites, has reduced. The most relevant reason for discovering novel secondary metabolites
is to circumvent the problem of resistant pathogens, which are no longer susceptible to the currently used drugs. Existence
of actinomycetes has been reported in the hitherto untapped marine ecosystem. Marine actinomycetes are efficient producers
of new secondary metabolites that show a range of biological activities including antibacterial, antifungal, anticancer, insecticidal
and enzyme inhibition. Bioactive compounds from marine actinomycetes possess distinct chemical structures that may form the
basis for synthesis of new drugs that could be used to combat resistant pathogens. 相似文献
95.
Lohray BB Lohray VB Srivastava BK Gupta S Solanki M Kapadnis P Takale V Pandya P 《Bioorganic & medicinal chemistry letters》2004,14(12):3139-3142
A number of substituted piperazinyl oxazolidinone derivatives have been synthesized and their antibacterial activities were evaluated by MIC determination. A systematic SAR was carried out to get highly potent oxazolidinone derivatives. 相似文献
96.
Fingerprinting method for phylogenetic classification and identification of microorganisms based on variation in 16S rRNA gene sequences 总被引:2,自引:0,他引:2
The paper describes a method for the classification and identification of microorganisms based on variations in 16S rRNA sequences. The 16S rRNA is one of the most conserved molecules within a cell. The nature of the variable and spacer regions has been found to be specific to a given organism. Thus, the method presented here can be very useful for the classification and identification of microorganisms for which very little information is available. To automate the method, a comprehensive computer program called FPMAP has been developed for the analysis of restriction fragment pattern data. The method involves the restriction digestion of genomic DNA, preferably using four-cutters that may recognize 6-9 sites within the 16S rDNA. The fragments are separated on a polyacrylamide gel along with a suitable marker, then transferred into a nylon membrane and hybridized with a radiolabeled 16S rDNA probe. After autoradiography, the fragment sizes are calculated, and the data are analyzed using the FPMAP software. We demonstrate that the method can be used for identification of strains of Streptomyces and mycobacteria. The software is available from our ftp site ftp:?imtech.chd.nic.in/pub/com/fpmap/unix/. 相似文献
97.
98.
Duffy KE Lamb RJ San Mateo LR Jordan JL Canziani G Brigham-Burke M Korteweg J Cunningham M Beck HS Carton J Giles-Komar J Duchala C Sarisky RT Mbow ML 《Cellular immunology》2007,248(2):103-114
Toll-like receptors are a family of pattern-recognition receptors that contribute to the innate immune response. Toll-like receptor 3 (TLR3) signals in response to foreign, endogenous and synthetic ligands including viral dsRNA, bacterial RNA, mitochondrial RNA, endogenous necrotic cell mRNA and the synthetic dsRNA analog, poly(I:C). We have generated a monoclonal antibody (mAb CNTO2424) that recognizes the extracellular domain (ECD) of human TLR3 in a conformation-dependent manner. CNTO2424 down-regulates poly(I:C)-induced production of IL-6, IL-8, MCP-1, RANTES, and IP-10 in human lung epithelial cells. In addition, mAb CNTO2424 was able to interfere with the known TLR3-dependent signaling pathways, namely NF-κB, IRF-3/ISRE, and p38 MAPK. The generation of this neutralizing anti-TLR3 mAb provides a unique tool to better understand TLR3 signaling and potential cross-talk between TLR3 and other molecules. 相似文献
99.
Arya SK Prusty AK Singh SP Solanki PR Pandey MK Datta M Malhotra BD 《Analytical biochemistry》2007,363(2):210-218
Cholesterol oxidase (ChOx) has been covalently immobilized onto two-dimensional self-assembled monolayer (SAM) of N-(2-aminoethyl)-3-aminopropyl-trimethoxysilane (AEAPTS) deposited on the indium-tin oxide (ITO) coated glass plates using N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide (EDC/NHS) chemistry. These ChO x/AEAPTS/ITO bioelectrodes are characterized using contact angle (CA) measurements, UV-visible spectroscopy, atomic force microscopy (AFM), electrochemical impedance technique, and Fourier transform infrared (FT-IR) technique. The covalently immobilized ChOx-modified AEAPTS bioelectrodes are used for the estimation of cholesterol in solution using UV-visible technique. These cholesterol sensing bioelectrodes show linearity as 50 to 500 mg/dl for cholesterol solution, detection limit as 25mg/dl, sensitivity as 4.499 x 10(-5) Abs (mg/dl)(-1), K(m) value as 58.137 mg/dl (1.5mM), apparent enzyme activity as 1.81 x 10(-3) U cm(-2), shelf life of approximately 10 weeks, and electrode reusability as 10 times. 相似文献
100.
Alasdair Cohen Yong Tao Qing Luo Gemei Zhong Jeff Romm John M. Colford Jr Isha Ray 《PloS one》2015,10(9)