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정금영,장성연,장성호,정규열 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-
Caprolactam is is a cyclic amide of caproic acid and is widely used to produce nylon-6 through ring-opening polymerization. An intracellular biosensor for caprolactam can facilitate high-throughput metabolic engineering of recombinant microbial strains. Because of the mixed production of caprolactam and valerolactam in the recombinant strain, a caprolactam biosensor should be highly specific for caprolactam. However, a highly specific caprolactam sensor has not been reported. Here, we developed a specific caprolactam riboswitch useful for monitoring the intracellular concentration of caprolactam. The riboswitch developed through coupled in vitro-in vivo selection responded specifically to caprolactam. Finally, caprolactam-dependent control of cell growth was demonstrated using the riboswitch. This riboswitch is a valuable tool for directed evolution of enzymes and pathway optimization for the microbial production of caprolactam.
정금영,신기영,정규열 한국공업화학회 2019 한국공업화학회 연구논문 초록집 Vol.2019 No.1
Microchip electrophoresis is very useful method for genetic analysis because of rapid analysis, small amount of sample/reagent consumption, and portability. Single strand conformation polymorphism (SSCP) analysis is one of promising genotyping method due to its simplicity and versatility. In SSCP analysis, the electrophoretic mobility variation comes from sequence-induced folded conformation difference of single strand DNA. Based on this principle, even a single base difference between DNA fragments being tested can be discriminated. However, the most critical limitation of SSCP analysis on microchip is low separation resolution resulting from short separation channel length. To develop high-resolution microchip SSCP analysis system, we investigated the relationship between the resolution of SSCP and the channel width. This analysis system was successfully applied to the various genetic analyses including food-borne pathogen detection and SNP marker detection.