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Novel Tools for Use in Bioluminescence Resonance Energy Transfer (BRET) Assays

Recent advances in imaging assays based on bioluminescence resonance energy transfer (BRET) have made it possible to study protein/protein interactions in living cells under physiological conditions. Here we describe protocols for these assays including relevant positive a ...

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Bioluminescence Imaging of Calcium Oscillations Inside Intracellular Organelles

Ca2+ oscillations inside intracellular organelles are important for regulation of functions such as gene expression at the nucleus, respiration at mitochondria or protein processing at the endoplasmic reticulum. Targeted aequorins are excellent calcium probes for subcell ...

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Bioluminescence Analysis of Smad-Dependent TGF- Signaling in Live Mice

TGF-β signaling via the Smad2/3 pathway has key roles in development and tissue homeostasis. Perturbations of the TGF-β signaling are involved in the pathogenesis of many human diseases, including cancer, fibrotic disorders, developmental defects, and neurodegeneration. To study ...

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Bioluminescent Imaging of MAPK Function with Intein-Mediated Reporter Gene Assay

For nondestructive analysis of chemical processes in living mammalian cells, here we show a new reporter gene assay for detecting Ras–Raf-1 interactions based on protein splicing of transcription factors with DnaE inteins. The protein splicing induces connection of a DNA-binding pr ...

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Analysis of ProteinProtein Interactions Using Bioluminescence Resonance Energy Transfer

Knowledge of how and when proteins interact in living cells is fundamental to our understanding of cellular biology, and bioluminescence resonance energy transfer (BRET) provides an increasingly popular mechanism for studying these interactions in real time. The technique util ...

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Quantitative In Vivo Imaging of Non-viral-Mediated Gene Expression and RNAi-Mediated Knockdown

Bioluminescent imaging (BLI) coupled with hydrodynamic (HD) dosing of luciferase-expressing plasmid DNA (pDNA) has proven to be a powerful method for quantitatively benchmarking non-viral gene expression in the liver. The expression of luciferase or knockdown of luciferase by RNA ...

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Real-Time Bioluminescence Imaging of Viral Pathogenesis

Mouse models are used commonly to study viral infection and define viral and host determinants of infection and disease morbidity. Conventional studies of viral infection in mice rely upon euthanizing cohorts of animals at multiple time points to identify sites of infection, quantify vi ...

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Bioluminescent Monitoring of In Vivo Colonization and Clearance Dynamics by Light-Emitting Bacteria

Bioluminescence is an excellent reporter system for analysing bacterial colonization and clearance dynamics in vivo. Many bacterial species have been rendered bioluminescent, allowing the sensitive detection of bacterial burden and metabolic activity in real-time and in si ...

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Noninvasive Bioluminescent Imaging of Infections

Traditional studies of viral and bacterial infection and pathogenesis have generally relied on animal models that require the sacrifice of infected animals to determine viral or bacterial distributions and titers. The recent application of the in vivo bioluminescence imaging (B ...

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Lipase and Phospholipase Biosensors: A Review

Recent advances in the field of biology, electronics, and nanotechnology have improved the development of biosensors. A biosensor is a device composed of a biological recognition element and a sensor element. Biosensor applications are becoming increasingly important in areas such ...

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Phospholipases in Food Industry: A Review

Mammal, plant, and mainly microbial phospholipases are continuously being studied, experimented, and some of them are even commercially available at industrial scale for food industry. This is because the use of phospholipases in the production of specific foods leads to attractive a ...

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Synthesis of Biodegradable Polymers Using Biocatalysis with Yarrowia lipolytica Lipase

Yarrowia lipolytica lipase (YLL) was used as catalyst in the enzymatic ring-opening polymerization (ROP) of ε-caprolactone. This low-cost solid-state lipase produces low-molecular-weight polyesters with unique multiphase morphology as determined by carbon-13 NMR. YLL att ...

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Lipases as Biocatalyst for Biodiesel Production

The global shortages of fossil fuels, significant increase in the price of crude oil, and increased environmental concerns have stimulated the rapid growth in biodiesel production. Biodiesel is generally produced through transesterification reaction catalyzed either chem ...

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Lipases in Green Chemistry: Acylation and Alcoholysis on Steroids and Nucleosides

In this article, we describe the application of lipases in acylation and alcoholysis reactions on steroids and nucleosides. In the field of steroids, a variety of acetyl and fatty acid derivatives of androstanes, pregnanes, and cholestanes have been prepared through lipase-catalyzed a ...

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Lipases as Catalysts in Synthesis of Fine Chemicals

The application of lipases as catalysts in the synthesis of an intermediate of alfuzosin and lapyrium chloride is described. In the first case, the one-pot procedure to obtain the intermediate involves the treatment of tetrahydrofuroic acid with ethanol in the presence of Candida antarc ...

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Lipase-Catalyzed Modification of Phenolic Antioxidants

The chemical acylation of natural antioxidants may improve their oxidative and thermal stability, as well as modify their hydrophile–lipophile balance (HLB). These processes are generally carried out under harsh conditions using strongly corrosive acids. In contrast, lipase- ...

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Lipases as Biocatalysts for the Synthesis of Structured Lipids

Structured lipids (SL) are broadly referred to as modified or synthetic oils and fats or lipids with functional or pharmaceutical applications. Some structured lipids, such as triglycerides that contain both long-chain (mainly essential) fatty acids and medium- or short-chain fatty ...

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Thermodynamical Methods for the Optimization of Lipase-Catalyzed Reactions

A basic insight on different thermodynamical strategies reported for the optimization of lipase-catalyzed reactions is presented. The significance of selecting the appropriate reaction media in order to enhance selectivity and operational stability of enzymes is discusse ...

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Rational Design of Immobilized Lipases and Phospholipases

Immobilization of lipases and phospholipases on, mainly, water insoluble carriers, helps in their economic reuse and in the development of continuous bioprocesses. Design of efficient lipases and phospholipases-immobilized system is rather a difficult task. A lot of research work ...

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Different Strategies for Hyperactivation of Lipase Biocatalysts

One important parameter for the application of lipase catalysts in chemical industries is the specific activity displayed towards natural or unnatural substrates. Different strategies to enhance the lipase activity have been described. The immobilization of lipases on hydrop ...

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