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排序方式: 共有52条查询结果,搜索用时 15 毫秒
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Mohamed I. Elzagheid Mikko Oivanen Karel D. Klika Bryan C. N. M. Jones Richard Cosstick Harri Lönnberg 《Nucleosides, nucleotides & nucleic acids》2013,32(9):2093-2108
Abstract The course of hydrolysis of 3′-deoxy-3′-thioinosylyl-(3′ → 5′)-uridine (IspU) has been followed by HPLC over a wide pH-range. Two reactions of the internucleosidic thiophosphate linkage compete: (i) cleavage yielding thioinosine monophosphates and uridine, and (ii) isomerization to the 2′,5′-isomer of IspU. Under very acidic conditions, even acid-catalyzed depurination of the inosine moiety is observed. The stability of the thiophosphate linkage and the mechanisms of its rupture are discussed. 相似文献
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Creation of genome-wide protein expression libraries using random activation of gene expression 总被引:8,自引:0,他引:8
Harrington JJ Sherf B Rundlett S Jackson PD Perry R Cain S Leventhal C Thornton M Ramachandran R Whittington J Lerner L Costanzo D McElligott K Boozer S Mays R Smith E Veloso N Klika A Hess J Cothren K Lo K Offenbacher J Danzig J Ducar M 《Nature biotechnology》2001,19(5):440-445
Here we report the use of random activation of gene expression (RAGE) to create genome-wide protein expression libraries. RAGE libraries containing only 5 x 10(6) individual clones were found to express every gene tested, including genes that are normally silent in the parent cell line. Furthermore, endogenous genes were activated at similar frequencies and expressed at similar levels within RAGE libraries created from multiple human cell lines, demonstrating that RAGE libraries are inherently normalized. Pools of RAGE clones were used to isolate 19,547 human gene clusters, approximately 53% of which were novel when tested against public databases of expressed sequence tag (EST) and complementary DNA (cDNA). Isolation of individual clones confirmed that the activated endogenous genes can be expressed at high levels to produce biologically active proteins. The properties of RAGE libraries and RAGE expression clones are well suited for a number of biotechnological applications including gene discovery, protein characterization, drug development, and protein manufacturing. 相似文献
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Petr Kovář Miroslav Pospíšil Petr Malý Zdeněk Klika Pavla Čapková Petra Horáková Marta Valášková 《Journal of molecular modeling》2009,15(11):1391-1396
The surface area of various types of montmorillonites (MMT) with different values of layer charge plays a very important role
in surface arrangement of methylene blue cations (MB). Photoluminescence measurements can be strongly or partially influenced
by this surface arrangement of cations. For these reasons and on the basis of our previous results, molecular simulations
were performed for various types of montmorillonites covered with methylene blue cations. Adsorption of methylene blue cations
on Na-Wyoming MMT surface is different from Ca-Cheto MMT. In the case of Wyoming with a lower layer charge, MB cations lie
parallel to the silicate layer for all investigated samples. On the other hand, Cheto surface is covered with a higher amount
of MB cations. The results obtained from molecular modeling indicate that MB lies parallel to low loading case and become
tilted with respect to layer for a higher loading. Moreover, a higher amount of MB cations covering the silicate layer are
much less energy-stable. A higher loading of MB cations leads to aggregates but at low loading MB cations degrade to monomers. 相似文献
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Koskela S Söderholm PP Ainasoja M Wennberg T Klika KD Ovcharenko VV Kylänlahti I Auerma T Yli-Kauhaluoma J Pihlaja K Vuorela PM Teeri TH 《Planta》2011,233(1):37-48
A previously isolated cDNA molecule from Gerbera hybrida (Asteraceae) codes for a new chalcone synthase-like polyketide synthase, 2-pyrone synthase (2PS). 2PS is able to synthesise 4-hydroxy-6-methyl-2-pyrone (triacetolactone), a putative precursor for gerberin and parasorboside, two abundant glucosides in gerbera. In this study, we show that gerbera plants transformed with the gene for 2PS in an antisense orientation and unable to synthesise gerberin and parasorboside are susceptible to Botrytis cinerea infection. In addition to the preformed glucosides, the transgenic plants also lack several compounds that are induced in control plants when infected with the mould. Some of these induced substances are effective in inhibiting fungal growth both in vitro and in vivo. Two of the phytoalexins were identified as the aglycones of gerberin and trans-parasorboside. The third phytoalexin is a rare coumarin, 4-hydroxy-5-methylcoumarin; however, it is typical of many plants of the sunflower family Asteraceae. The coumarin cannot be structurally derived from either gerberin or parasorboside, but may be derived from a related polyketide intermediate. 相似文献
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