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近 2 0年来 ,人工核酸切割试剂的研究一直是化学、生物化学和分子生物学中最为活跃的前沿领域之一[1,2 ] .人工核酸切割试剂可以在足迹技术和核酸高级结构的研究中用作高分辨率的化学探针 ,还可以用于合成定点切割试剂[3] .后者又被称为人工工具酶 ,是一种非常重要的分子生物学工具 ,在疾病的基因治疗、反义 PCR技术等领域中都具有重要的应用 .人工核酸切割试剂的切割机理主要有自由基机理和磷酸酯水解机理两大类 .相对于自由基机理 ,水解机理具有许多优点 ,使得水解型切割试剂具有更为广泛的应用 .对于 DNA,目前文献报道的水解型人工切… 相似文献
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具有DNA切割功能的新型多聚酰胺/丝组缀合物 总被引:1,自引:0,他引:1
为得到具有核酸切割功能的人工核酸酶, 设计合成了一种新型多聚酰胺/丝组缀合物, 并研究了其DNA切割活性. 合成的目标化合物在pH=6.0的BR缓冲溶液中对pBR322 DNA切割活性的初步实验结果表明, 于37 ℃保温6 h后, pBR322 DNA基本上被完全从Form Ⅰ切割为Form Ⅱ, 保温36 h后, pBR322 DNA几乎被切割完全. 相似文献
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采用1-(3-二甲氨基丙基)-3-乙基碳二亚胺方法合成氨基甲酸乙酯(Ethyl carbamate,EC)新型人工抗原.将衰减全反射红外光谱法(ATR-FTIR)应用于人工合成抗原的表征分析,并结合荧光光谱分析EC人工抗原的偶联效果以及载体蛋白质分子的二级结构变化;通过质谱结合紫外光谱、电泳方法进行人工抗原的系统表征,计算新型人工抗原中半抗原分子与载体蛋白质分子的偶联比.结果表明:合成路线合理,成功获得了氨基甲酸乙酯新型人工抗原.人工抗原分子的α-螺旋、β-折叠和β-转角结构与载体蛋白质分子相比含量发生变化,人工抗原的荧光相图满足线性型态变迁关系,符合“二态模型”.氨基甲酸乙酯人工抗原分析表征的红外衰减全反射方法、基质辅助激光解析飞行时间质谱法获得的检测结果与其它光谱方法、电泳方法表征结果一致,获得人工抗原的偶联比为15∶1~19∶1,EC新型人工抗原免疫小鼠抗血清的效价为1∶25600. 相似文献
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丝组二肽对DNA的切割作用的研究 总被引:2,自引:2,他引:0
近20年来,人工核酸切割试剂的研究一直是生物化学中最为活跃的前沿领域之一,研究人工核酸切割试剂的主要目的是合成定点切割试剂,后者是一种重要的分子生物学工具,在疾病的基团治疗、反义PCR技术等领域中有着重要的应用价值。此外,人工核酸切割试剂还可以在足迹技术和核酸高级结构的研究中用作高分辨率的化学探针。 相似文献
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Valerian T. D'Souza 《Supramolecular chemistry》2013,25(3):221-229
The magical powers of enzymes have been attributed to their ability to bind specific substrates and catalyze reactions of the bound substrate. Artificial enzymes synthetically mimic the binding and the catalytic site to produce molecules that are not only smaller in size but also potentially have similar activity to the real enzymes. The main objective of our research is to create artificial redox enzymes by using cyclodextrins as binding sites and attaching flavin derivatives as the catalytic site. We have developed a strategy to attach a catalytic site to cyclodextrin exclusively at the 2-, 3- or the 6-position. The evaluation of the artificial enzyme in which flavin is attached to the 2-position gives a 647-fold acceleration factor. Although this is modest compared to those of real enzymes (which can have acceleration factors of a trillion), the artificial enzymes allow us to understand the elements that contribute to the incredible catalytic power of enzymes. 相似文献
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Rates of phosphotriester bond formation and amounts of sulphonation are compared for three popular coupling agents used in oligodeoxyribonucleotide synthesis by the phosphotriester approach. 相似文献
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Hajime Karatani 《Analytical sciences》2007,23(6):747-750
Anodic oxidation of oligodeoxyribonucleotide in an alkaline aqueous medium containing tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)3(2+)) was shown to cause luminescence around +1.3 V (vs. Ag/AgCl) with a maximal intensity at approximately 600 nm, possibly originating from Ru(bpy)3(2+) in the d-pi* triplet state. A pivotal initial stage in the light production path was postulated to be the anodic oxidation of 2-deoxyribose residue. This reaction seems to be available for the determination of sub-micromol dm(-3) levels of oligodeoxyribonucleotide. 相似文献
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Chen Yaoxia Zhang Wenwen Ding Yinghao Liang Chunhui Shi Yang Hu Zhi-Wen Wang Ling Yang Zhimou 《中国科学:化学(英文版)》2021,64(9):1554-1559
The creation of artificial enzymes to mimic natural enzymes remains a great challenge owing to the complexity of the structural arrangement of the essential amino acids in catalytic centers. In this study, we used the phosphatase-based enzyme-instructed self-assembly(EISA) to supervise artificial esterases' final structures and catalytic activities. We reported that peptide precursors containing different phosphorylation sites could preorganize into alternated nanostructures and undergo dephosphorylation in the presence of alkaline phosphatase(ALP) with variation in kinetic and thermodynamic profiles. Although identical self-assembly compositions were formed after dephosphorylation, precursors with more enhanced preorganized states tended to better promote ALP dephosphorylation, facilitate further self-assembly, and strengthen the catalytic activities of the final assemblies. We envisioned that our strategy would be useful for further construction and manipulation of various artificial enzymes with superior catalytic activities. 相似文献
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Dr. Sarah Lopez Dr. Caroline Marchi-Delapierre Dr. Christine Cavazza Dr. Stéphane Ménage 《Chemistry (Weinheim an der Bergstrasse, Germany)》2020,26(70):16633-16638
Performing a heterogeneous catalysis with proteins is still a challenge. Herein, we demonstrate the importance of cross-linked crystals for sulfoxide oxidation by an artificial enzyme. The biohybrid consists of the insertion of an iron complex into a NikA protein crystal. The heterogeneous catalysts displays a better efficiency-with higher reaction kinetics, a better stability and expand the substrate scope compared to its solution counterpart. Designing crystalline artificial enzymes represents a good alternative to soluble or supported enzymes for the future of synthetic biology. 相似文献
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Several peptide nucleic acid based artificial nucleases (PNAzymes) are designed to create a bulge in the target RNA, which is a short model of the leukemia related bcr/abl mRNA. The target RNA is cleaved by the PNAzymes with a half-life of down to 11 h (using a 1 : 1 ratio of PNA-conjugate to target) and only upon base-pairing with the substrate. The PNA based systems are also shown to act in a catalytic fashion with turnover of substrate and are thus the first reported peptide nucleic acid based artificial RNA-cleaving enzymes. 相似文献
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P. E. Vorobjev I. A. Pyshnaya E. Wickstrom V. F. Zarytova 《Russian Chemical Bulletin》2002,51(7):1187-1189
It was demonstarted for the first time that RNA can be subjected to site-specific oxidative cleavage induced by the glycopeptide antibiotic bleomycin A5 (Blm) covalently linked to the 3"-terminus of an oligodeoxyribonucleotide through two, three, or four residues of hexaethylene glycol phosphate (p-heg)
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. The oligonucleotide conjugate with bleomycin forms an imperfect complementary complex with the RNA to be cleaved (5"-prCGGAGUUGGAAAACAAUGAAAAGGCCCCCA/Blm-(p-heg)
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-3"-pdGCCTCACCTTTTGTTA). The cleavage occurs at the only nucleotide residue (U) preceding a one-nucleotide bulge in the RNA chain, which is formed due to imperfect complementarity between the oligodeoxyribonucleotide and the RNA to be cleaved. 相似文献
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For the detection of deletion polymorphisms, two pyrene moieties are tethered to an oligodeoxyribonucleotide (ODN) on both sides of the intervening base; one- and two-base deletions can be selectively detected by the strength of the excimer emission. 相似文献
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Weida Tong Hongping Ye Ding Rong Valerian T. D'Souza 《Journal of computational chemistry》1992,13(5):614-621
A computational chemistry study of the artificial redox enzyme synthesized by covalently attaching flavin to cyclodextrins explains some of its properties. Calculations indicate that the flavin moiety covalently attached to cyclodextrin is not within the cavity of cyclodextrin. This result is consistent with the UV-vis spectrum of the artificial enzyme. The calculations also indicate hydrogen bonds formed between the carbonyl groups of the catalytic functionality and the hydroxyl groups of cyclodextrin play a role in their most stable conformation. This explains the observed overall stability of these artificial enzymes compared to riboflavin. Electrostatic energies and solvation energies play a major role in the stability of the hosts and the orientation of guests included within the artificial enzymes. The rates of oxidation of various thiols catalyzed by the artificial enzyme can be explained by the relative distances between the sulfur atom of the substrates and C(4a) of the flavin moiety. 相似文献