The influence of high speed swimsuits on the lactate curve test among competitive swimmers

departmental bulletin pape

A Fluorescent Split Aptamer for Visualizing RNA–RNA Assembly <i>In Vivo</i>

RNA–RNA assembly governs key biological processes and is a powerful tool for engineering synthetic genetic circuits. Characterizing RNA assembly in living cells often involves monitoring fluorescent reporter proteins, which are at best indirect measures of underlying RNA–RNA hybridization events and are subject to additional temporal and load constraints associated with translation and activation of reporter proteins. In contrast, RNA aptamers that sequester small molecule dyes and activate their fluorescence are increasingly utilized in genetically encoded strategies to report on RNA-level events. Split-aptamer systems have been rationally designed to generate signal upon hybridization of two or more discrete RNA transcripts, but none directly function when expressed <i>in vivo</i>. We reasoned that the improved physiological properties of the Broccoli aptamer enable construction of a split-aptamer system that could function in living cells. Here we present the Split-Broccoli system, in which self-assembly is nucleated by a thermostable, three-way junction RNA architecture and fluorescence activation requires both strands. Functional assembly of the system approximately follows second-order kinetics <i>in vitro</i> and improves when cotranscribed, rather than when assembled from purified components. <i>Split-Broccoli</i> fluorescence is digital <i>in vivo</i> and retains functional modularity when fused to RNAs that regulate circuit function through RNA–RNA hybridization, as demonstrated with an RNA Toehold switch. <i>Split-Broccoli</i> represents the first functional split-aptamer system to operate <i>in vivo</i>. It offers a genetically encoded and nondestructive platform to monitor and exploit RNA–RNA hybridization, whether as an all-RNA, stand-alone AND gate or as a tool for monitoring assembly of RNA–RNA hybrids

Selective Inactivation of Functional RNAs by Ribozyme-Catalyzed Covalent Modification

The diverse functions of RNA provide numerous opportunities for programming biological circuits. We describe a new strategy that uses ribozyme K28min to covalently tag a specific nucleobase within an RNA or DNA target strand to regulate and selectively inactivate those nucleic acids. K28min variants with appropriately reprogrammed internal guide sequences efficiently tagged multiple sites from an mRNA and from aptamer and ribozyme targets. Upon covalent modification by the corresponding K28min variant, an ATP-binding aptamer lost all affinity for ATP, and the fluorogenic Mango aptamer lost its ability to activate fluorescence of its dye ligand. Modifying a hammerhead ribozyme near the catalytic core led to loss of almost all of its substrate-cleaving activity, but modifying the same hammerhead ribozyme within a tertiary stabilizing element that reduces magnesium dependence only impaired substrate cleavage at low magnesium concentration. Thus, ribozyme-mediated covalent modification can be used both to selectively inactivate and to fine-tune the activities of targeted functional RNAs, analogous to the effects of post-translational modifications of proteins. Ribozyme-catalyzed covalent modification could therefore be developed to regulate nucleic acids components of synthetic and natural circuits

Study on the perfection of in situ P-injection synthesis LEC-InP single crystals

Undoped, S-doped and Fe-doped InP crystals with diameter up to 4-inch have been pulled in drop 10 0 drop -direction under P-rich condition by a rapid P-injection in situ synthesis liquid encapsulated Czochralski (LEC) method. High speed photoluminescence mapping, etch-pit density (EPD) mapping and scanning electron microscopy have been used to characterize the samples of the single crystal ingots. Dislocations and electrical homogeneity of these samples are investigated and compared. By controlling the thermal field and the solid-liquid interface shape, 4-inch low-EPD InP single crystals have been successfully grown by the rapid P-injection synthesis LEC method. The EPD across the wafer of the ingots is less than 5 x 10(4) cm(-2). Cluster defects with a pore center are observed in the P-rich LEC grown InP ingots. These defects are distributed irregularly on a wafer and are surrounded by a high concentration of dislocations. The uniformity of the PL intensity across the wafer is influenced by these defects. (C) 2004 Elsevier B.V. All rights reserved