BackgroundTliA is a thermostable lipase secreted by the type 1 secretion system (T1SS) of Pseudomonas fluorescens. The secretion is promoted by its secretion/chaperone domain located near the C-terminus, which is composed mainly of four Repeat-in-Toxin (RTX) repeats. In order to identify the minimal region of TliA responsible for its secretion, five different copies of the secretion/chaperone domain, each involving truncated N-terminal residues and a common C-terminus, were acquired and named as lipase ABC transporter recognition domains (LARDs). Each LARD was fused to epidermal growth factor (EGF) or green fluorescent protein (GFP), and the secretion of EGF-LARD or GFP-LARD fusion proteins was assessed in Escherichia coli with ABC transporter.ResultsAmong the fusion proteins, GFP or EGF with 105-residue LARD3 was most efficiently secreted. In addition, GFP-LARD3 emitted wild type GFP fluorescence. Structurally, LARD3 had the 4 RTX repeats exposed at the N-terminus, while other LARDs had additional residues prior to them or missed some of the RTX repeats. LARD3 was both necessary and sufficient for efficient secretion and maintenance of GFP fluorescence in E. coli, which was also confirmed in P. fluorescens and P. fluorescens ▵tliA, a knock-out mutant of tliA.ConclusionLARD3 was a potent secretion signal in T1SS for its fusion flanking RTX motif, which enhanced secretion and preserved the fluorescence of GFP. LARD3-mediated secretion in E. coli or P. fluorescens will enable the development of enhanced protein manufacturing factory and recombinant microbe secreting protein of interest in situ.
Digital video content via broadcast television, Internet and other content distribution networks provide limited interaction for fitness and wellness activities. The content delivery is one-way only and provides no personalization to the pace, programming and progress of the user's exercise routine. Furthermore, the content is to be viewed only on a screen which makes it awkward and incompatible with full-body activities such as yoga, pilates and T'ai chi. We present CloudMat, a system for context-aware personalization of fitness content with cloudenabled connected surfaces. CloudMat provides real-time closedloop feedback between the state of the user on the physical mat and the state of the content in the cloud service. Content is tagged with actuation signals where events are delegated from the screen to display on an electroluminescent lighting layer on the mat, which provides spatial guidance to the end-user. Through the sensor-layer embedded in the mat, the physical interface captures the pose and timing of the user activity and relays it to the Context-aware Personalization cloud service. This service coordinates sensing and actuation between the content stream and mat by generating pose templates and metadata files about the exercise routine to be delivered to the user. Through this interactive process between the physical mat and the content service, the feedback provided by the user performing the routine continuously adapts the pace and programming to maintain the desired user experience. We demonstrate the utility of the system and evaluate the system performance with a case study on interactive yoga. Abstract-Digital video content via broadcast television, Internet and other content distribution networks provide limited interaction for fitness and wellness activities. The content delivery is one-way only and provides no personalization to the pace, programming and progress of the user's exercise routine. Furthermore, the content is to be viewed only on a screen which makes it awkward and incompatible with full-body activities such as yoga, pilates and T'ai chi. We present CloudMat, a system for context-aware personalization of fitness content with cloudenabled connected surfaces. CloudMat provides real-time closedloop feedback between the state of the user on the physical mat and the state of the content in the cloud service. Content is tagged with actuation signals where events are delegated from the screen to display on an electroluminescent lighting layer on the mat, which provides spatial guidance to the end-user. Through the sensor-layer embedded in the mat, the physical interface captures the pose and timing of the user activity and relays it to the Context-aware Personalization cloud service. This service coordinates sensing and actuation between the content stream and mat by generating pose templates and metadata files about the exercise routine to be delivered to the user. Through this interactive process between the physical mat and the content service, the feedback provided by the ...
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