Rapid generation of incremental truncation libraries for protein engineering using α-phosphothioate nucleotides

Incremental truncation for the creation of hybrid enzymes (ITCHY) is a novel tool for the generation of combinatorial libraries of hybrid proteins independent of DNA sequence homology. We herein report a fundamentally different methodology for creating incremental truncation libraries using nucleotide triphosphate analogs. Central to the method is the polymerase catalyzed, low frequency, random incorporation of α-phosphothioate dNTPs into the region of DNA targeted for truncation. The resulting phosphothioate internucleotide linkages are resistant to 3′→5′ exonuclease hydrolysis, rendering the target DNA resistant to degradation in a subsequent exonuclease III treatment. From an experimental perspective the protocol reported here to create incremental truncation libraries is simpler and less time consuming than previous approaches by combining the two gene fragments in a single vector and eliminating additional purification steps. As proof of principle, an incremental truncation library of fusions between the N-terminal fragment of Escherichia coli glycinamide ribonucleotide formyltransferase (PurN) and the C-terminal fragment of human glycinamide ribonucleotide formyltransferase (hGART) was prepared and successfully tested for functional hybrids in an auxotrophic E.coli host strain. Multiple active hybrid enzymes were identified, including ones fused in regions of low sequence homology.

JavaVis: An Integrated Computer Vision Library for Teaching Computer Vision

In this article, we present a new framework oriented to teach Computer Vision related subjects called JavaVis. It is a computer vision library divided in three main areas: 2D package is featured for classical computer vision processing; 3D package, which includes a complete 3D geometric toolset, is used for 3D vision computing; Desktop package comprises a tool for graphic designing and testing of new algorithms. JavaVis is designed to be easy to use, both for launching and testing existing algorithms and for developing new ones.

Experiences Using an Open Source Sofware Library to Teach Computer Vision Subjects

Machine vision is an important subject in computer science and engineering degrees. For laboratory experimentation, it is desirable to have a complete and easy-to-use tool. In this work we present a Java library, oriented to teaching computer vision. We have designed and built the library from the scratch with enfasis on readability and understanding rather than on efficiency. However, the library can also be used for research purposes. JavaVis is an open source Java library, oriented to the teaching of Computer Vision. It consists of a framework with several features that meet its demands. It has been designed to be easy to use: the user does not have to deal with internal structures or graphical interface, and should the student need to add a new algorithm it can be done simply enough. Once we sketch the library, we focus on the experience the student gets using this library in several computer vision courses. Our main goal is to find out whether the students understand what they are doing, that is, find out how much the library helps the student in grasping the basic concepts of computer vision. In the last four years we have conducted surveys to assess how much the students have improved their skills by using this library.

Workshops in “Construction of Structures I”: A new teaching methodology in the Building Engineering Degree

Both the current economic situation in the construction sector and the continuous normative changes in the building area imply the use of new methodologies to enhance students’ competences in the degree of Building Engineer. The aim of this paper is to present, analyse and discuss the development of constructive workshops as a new teaching methodology used in the subject of Construction of Structures I at the University of Alicante to complement the constructive and technical knowledge acquired by our students and to enhance their communicative and representation skills essential for their professional practice in the future. The used methodology is based on the development of three-dimensional construction details (in groups of 3 or 4 students) to be shown in two A1-pannels exposed in the corridors of the Polytechnic School. Thus, students’ work approaches constructive problems in a global way by discussing simultaneously with teachers and other groups about the most suitable solution on each case. This contribution has multiperspective results and improves criticism of students in different areas, encouraging new learning strategies and active participation. What is more, on-line information and web applications have been used to prepare and organize this kind of workshops, allowing students to use new technologies as a complementary learning methodology. In conclusion, the use of these new workshops in the Degree of Building Engineer stimulates an interactive class versus a traditional lecture where the participative groups´ attitude and the development of oral presentations dissolve the traditional boundaries regarding public communication skills of the students in the Degree.

Students at University of Michigan do research in the electrical engineering laboratory in the College of Engineering

Shown recording data are, left to right, John P. Vajda, Milwaukee WI, and James A. Lovell, Grand Rapids, MI. graduate students in electrical engineering.

West Engineering Annex

View from the northeast. Old carpenter's shop is on the right next to the Annex which was built to replace it. On verso: Engineering shops

West Engineering Annex

Profs. J.B. Davis & Mortimer Cooley, architects? Engineering Shops from 1885-1923. Later used as West Engineering Annex 1923. Architecture Dept. in east wing 1923-1927; Survey Dept. in east wing 1927; Auto Lab in west wing; part of Auto Lab destroyed by fire 1937; rest demolished 1956.

West Engineering Annex

Addition comprised of a central section with tower and a one story wing on the west. Engineering shops. Several people in image.

West Engineering Annex

Profs. J.B. Davis & Mortimer Cooley, architects? Engineering Shops from 1885-1923. Later used as West Engineering Annex 1923. Architecture Dept. in east wing 1923-1927; Survey Dept. in east wing 1927; Auto Lab in west wing; part of Auto Lab destroyed by fire 1937; rest demolished 1956. On verso: Engineering Laboratory. Female person in image.

Engineering Annex

Profs. J.B. Davis & Mortimer Cooley, architects? Engineering Shops from 1885-1923. Later used as West Engineering Annex 1923. Architecture Dept. in east wing 1923-1927; Survey Dept. in east wing 1927; Auto Lab in west wing; part of Auto Lab destroyed by fire 1937; rest demolished 1956. In 1918, clock and chimes were removed from the Old General Library and installed in the tower of the Engineering Annex. Old General Library was torn down in 1918.

Engineering Laboratory - U of M

Photograph of drawing by Benno Rohnert. On verso: "Center- First Engineering Bldg. Brick & frame 1881-82. Right-Abandoned campus carpenter shop moved against west side 1883 and used for wood shop. Left-First unit of 3 story brick building started 1885" Original two buildings removed in 1888.

Electrical Engineering Laboratory

l-r John P. Vajda, James A. Lovell grad students