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Enzymatic production of RNAi libraries from cDNAs   总被引:30,自引:0,他引:30  
RNA interference (RNAi) induced by small interfering (siRNA) or short hairpin RNA (shRNA) is an important research approach in mammalian genetics. Here we describe a technology called enzymatic production of RNAi library (EPRIL) by which cDNAs are converted by a sequence of enzymatic treatments into an RNAi library consisting of a vast array of different shRNA expression constructs. We applied EPRIL to a single cDNA source and prepared an RNAi library consisting of shRNA constructs with various RNAi efficiencies. High-throughput screening allowed us to rapidly identify the best shRNA constructs from the library. We also describe a new selection scheme using the thymidine kinase gene for obtaining efficient shRNA constructs. Furthermore, we show that EPRIL can be applied to constructing an RNAi library from a cDNA library, providing a basis for future whole-genome phenotypic screening of genes.  相似文献   
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Formation of myelin sheaths by Schwann cells (SCs) enables rapid and efficient transmission of action potentials in peripheral axons, and disruption of myelination results in disorders that involve decreased sensory and motor functions. Given that construction of SC myelin requires high levels of lipid and protein synthesis, mitochondria, which are pivotal in cellular metabolism, may be potential regulators of the formation and maintenance of SC myelin. Supporting this notion, abnormal mitochondria are found in SCs of neuropathic peripheral nerves in both human patients and the relevant animal models. However, evidence for the importance of SC mitochondria in myelination has been limited, until recently. Several studies have recently used genetic approaches that allow SC-specific ablation of mitochondrial metabolic activity in living animals to show the critical roles of SC mitochondria in the development and maintenance of peripheral nerve axons. Here, we review current knowledge about the involvement of SC mitochondria in the formation and dysfunction of myelinated axons in the peripheral nervous system.  相似文献   
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Zusammenfassung Die Wirkung elektrischer Reizung einerseits vagal, andererseits transmural auf isolierte Muskelpräparate vom Hühnervormagen wurde untersucht und gefunden, dass eine nach Aufhören der Reizung beobachtete Nachkontraktion nicht als «rebound» zu betrachten ist, sondern cholinergische Züge zeigt.  相似文献   
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引经据典是李善注的最大特点,与五臣注比较,被认为没有“释义“,《六臣注文选》“善曰言“有数百处,这些注是李善表现自己见解的阐述。李善注解释的内容与五臣注解释的内容有关系,以《六臣注文选》卷二十二谢灵运作品的解释为中心,研讨两者的思想观点。  相似文献   
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S Oikawa  M Imai  A Ueno  S Tanaka  T Noguchi  H Nakazato  K Kangawa  A Fukuda  H Matsuo 《Nature》1984,309(5970):724-726
Recent identification of natriuretic-diuretic activity in peptides isolated from human and rat atrial tissue implicates them in the control of extracellular fluid volume and electrolytic homeostasis. The presence of multiple forms of the peptides ranging from 3,000 to 13,000 molecular weight (MW) suggests they may all derive from the same precursor. The established amino acid sequence of alpha-human atrial natriuretic polypeptide (alpha- hANP ), a 28-residue peptide with potent natriuretic activity, provided the means to elucidate the structure of the precursor for alpha- hANP and the gene encoding it. Here we report the cloning and sequence analysis of the cDNA of human atrial mRNA encoding a precursor of alpha- hANP . The cDNA encodes gamma-human atrial natriuretic polypeptide (gamma- hANP ) of 13,000 MW, whose C-terminal 28 amino acid residues may be processed as alpha- hANP .  相似文献   
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Thomas L  Hayashi M  Jiang X  Moriya R  Rettner C  Parkin SS 《Nature》2006,443(7108):197-200
Magnetic domain walls, in which the magnetization direction varies continuously from one direction to another, have long been objects of considerable interest. New concepts for devices based on such domain walls are made possible by the direct manipulation of the walls using spin-polarized electrical current through the phenomenon of spin momentum transfer. Most experiments to date have considered the current-driven motion of domain walls under quasi-static conditions, whereas for technological applications, the walls must be moved on much shorter timescales. Here we show that the motion of domain walls under nanosecond-long current pulses is surprisingly sensitive to the pulse length. In particular, we find that the probability of dislodging a domain wall, confined to a pinning site in a permalloy nanowire, oscillates with the length of the current pulse, with a period of just a few nanoseconds. Using an analytical model and micromagnetic simulations, we show that this behaviour is connected to a current-induced oscillatory motion of the domain wall. The period is determined by the wall's mass and the slope of the confining potential. When the current is turned off during phases of the domain wall motion when it has enough momentum, the domain wall is driven out of the confining potential in the opposite direction to the flow of spin angular momentum. This dynamic amplification effect could be exploited in magnetic nanodevices based on domain wall motion.  相似文献   
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