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korea research institute of bioscience and biotechnology

3 ARTICLES PUBLISHED IN JoVE

Biology

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Haseong Kim¹, Kil Koang Kwon¹, Wonjae Seong¹, Seung-Goo Lee^1,2

¹Synthetic Biology & Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology, ²Biosystems and Bioengineering Program, University of Science and Technology, Daejeon, South Korea

This work presents a method of high-throughput screening using a universal genetic enzyme screening system that can be theoretically applied to over 200 enzymes. Here, the single screening system identifies three different enzymes (lipase, cellulase, and alkaline phosphatase) by simply changing the substrate used (p-nitrophenyl acetate, p-nitrophenyl-β-D-cellobioside, and phenyl phosphate).

Biochemistry

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Haseong Kim^*1, Gui Hwan Han^*1, Yaoyao Fu¹, Jongsik Gam², Seung Goo Lee^1,3

¹Synthetic Biology & Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology, ²College of Interdisciplinary & Creative Studies, Konyang University, ³Biosystems and Bioengineering Program, University of Science and Technology

This protocol describes the rapid and highly sensitive quantification of Förster resonance energy transfer (FRET) sensor data using a custom-made portable FRET analyzer. The device was used to detect maltose within a critical temperature range that maximizes detection sensitivity, enabling practical and efficient assessment of sugar content.

Genetics

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow

Gajendra W. Suryawanshi^*1,2, Song Xu^*3, Yiming Xie¹, Tom Chou³, Namshin Kim⁴, Irvin S. Y. Chen^1,5, Sanggu Kim⁶

¹UCLA AIDS Institute, University of California at Los Angeles (UCLA), ²Department of Microbiology, Immunology, & Molecular Genetics, University of California at Los Angeles (UCLA), ³Departments of Biomathematics and Mathematics, University of California at Los Angeles (UCLA), ⁴Personalized Genomic Medicine Research Center, Division of Strategic Research Groups, Korea Research Institute of Bioscience and Biotechnology, ⁵Department of Medicine, University of California at Los Angeles (UCLA), ⁶Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University (OSU)

This manuscript describes the experimental procedure and software analysis for a bidirectional integration site assay that can simultaneously analyze upstream and downstream vector-host junction DNA. Bidirectional PCR products can be used for any downstream sequencing platform. The resulting data are useful for a high-throughput, quantitative comparison of integrated DNA targets.