Efficient Linear dsDNA Tagging Using Deoxyuridine Excision**

Site-specific strategies for exchanging segments of dsDNA are important for DNA library construction and molecular tagging. Deoxyuridine (dU) excision is an approach for generating 3’ ssDNA overhangs in gene assembly and molecular cloning procedures. Unlike approaches that use a multi-base pair motif to specify a DNA cut site, dU excision requires only a dT→dU substitution. Consequently, excision sites can be embedded in biologically active DNA sequences by placing dU substitutions at non-perturbative positions. In this work, I describe a molecular tagging method that uses dU excision to exchange a segment of a dsDNA strand with a long synthetic oligonucleotide. The core workflow of this method, called deoxyUridine eXcision-tagging (dUX-tagging), is an efficient one-pot reaction: strategically positioned dU nucleotides are excised from dsDNA to generate a 3’ overhang so that additional sequence can be appended by annealing and ligating a tagging oligonucleotide. The tagged DNA is then processed by one of two procedures to fill the 5’ overhang and remove excess tagging oligo. To facilitate its widespread use, all dUX-tagging procedures exclusively use commercially available reagents. As a result, dUX-tagging is a concise and easily implemented approach for high-efficiency linear dsDNA tagging.     

Robot–human rescue teams: a user requirements analysis

Time-critical operations of equipment in emergencies require efficient human–machine interaction. In order to evaluate and rank the factors affecting search and rescue activities in emergencies, a requirements analysis among this professional community has been performed. The analyses presented here focus on cooperating robot–human teams. They have been performed on the basis of questionnaires and personal interviews with professionals in search and rescue from Germany, Finland and the Czech Republic. This includes professional fire fighters, plant fire brigades (nuclear power plants, airports, chemical factories), governmental disaster relief organizations and the military. The analyses identify the end-user requirements and provide guidelines for the development of rescue systems. On this basis, a system design for human–robot teams has been derived, with particular emphasis on telerobotics interface implemenation.     

Modular behavior-based control for team humanoids

When developing teams of humanoids — rather than an individual humanoid — there are a number of issues that become important to consider, including robustness, scalability, versatility, and also development and production costs. Therefore, we used a modern approach to AI that puts emphasis on the balance between control, electronic hardware, material, sensory system and energy in order to develop the team of Viki humanoid robots. In contrast to the top-down approach of equipping a humanoid with as many sensors, motors, power, etc., as possible, we developed a bottom-up approach to the construction of humanoids, regarding both hardware and software (modular behaviorbased control). The approach is shown with the development of the Viki humanoid team that won the RoboCup Humanoids Free Style World Championship 2002. In this paper, we focus on the main result of the behavior-based architecture with many layers of behaviors at different levels, which make it easy for both engineers to design new behaviors and for end-users to develop humanoid behaviors at different levels of complexity, dependent on the competencies of the individual end-user. With this architecture, it becomes possible to develop simple user interfaces with a user-guided implementation of our modular behavior-based approach, in order to allow any user to design performances with the humanoid robots.     

Development of the assistive mobile robot system: AMOS—to aid in the daily life of the physically handicapped

It can be exhausting, both physically and emotionally, to assist physically handicapped persons. Therefore, an assistive mobile system called AMOS (Assistive MObile robot System) has been developed to help alleviate this burden. The purpose of AMOS is to pick up and transport daily use objects, placing them in a designated indoor location semi-autonomously. AMOS consists of a self-contained mobile robot body, a user interface with a touch-panel and a computer network (Ethernet LAN, Internet). The user interacts with the mobile robot through a Web browser connected to a computer network, allowing for communication anytime, from anywhere and by anyone. This mode provides a simple way for communicating with and determining the status of the robot. Experiments were performed to verify the successful operation of AMOS. Although the system performed as designed, it would prove useful to the extend service area of the robot through communication mechanisms and the user interface.     

CRISPR/Cas9-RNA interference system for combinatorial metabolic engineering of Saccharomyces cerevisiae

The yeast Saccharomyces cerevisiae is widely used in industrial biotechnology for the production of fuels, chemicals, food ingredients, food and beverages, and pharmaceuticals. To obtain high-performing strains for such bioprocesses, it is often necessary to test tens or even hundreds of metabolic engineering targets, preferably in combinations, to account for synergistic and antagonistic effects. Here, we present a method that allows simultaneous perturbation of multiple selected genetic targets by combining the advantage of CRISPR/Cas9, in vivo recombination, USER assembly and RNA interference. CRISPR/Cas9 introduces a double-strand break in a specific genomic region, where multiexpression constructs combined with the knockdown constructs are simultaneously integrated by homologous recombination. We show the applicability of the method by improving cis,cis-muconic acid production in S. cerevisiae through simultaneous manipulation of several metabolic engineering targets. The method can accelerate metabolic engineering efforts for the construction of future cell factories.     

多语言用户界面的研究与实现

提出了一种适应多用户界面的工具—多语言转换器 ,详细论述了多语言转换器的实现技术 ,并对该技术的作用进行了分析 ,最后把它应用于软件开发中