원문정보
초록
영어
Recently, various secondary devices have been developed to detect the odorant chemical by using olfactory receptor as a primary transducer.
Detection of odorant at very low concentration has become a possibility by these devices. But currently there is no device that shows the odorant binding to the olfactory receptors with a fluorescence imaging. In this research, a cAMP response element reporter gene system was used for odorant screening. When odorant molecules bind to the olfactory receptor, the cAMP level was increased and initiated the phosphorylation of the CREB which binds to CRE promoter to induce the expression of ZsGreen as a reporter. For the high-throughput screening, PEG microwell was fabricated on the glass and reverse transfection technique was also used. Plasmid vector containing olfactory receptor gene and lipofectamin were mixed with gelatin solution and spotted on the PEG microwell.
HEK293 cells were seeded on the glass substrate and olfactory receptors were expressed in 48 hours. The odorant binding event was detected by fluorescence of ZsGreen as a reporter.