Development and Application of a Bioluminescent Biosening System

• Main Authors: Liao, Yuh-Shyan, 廖育賢
• Other Authors: Chien-Yuan Chen
• Format: Others
• Language: zh-TW
• Published: 1997
• Online Access: http://ndltd.ncl.edu.tw/handle/04177803317494064022

碩士 === 國立臺灣大學 === 農業化學系 === 85 === Luminescence is a generic term that covers a range of process which produce light. This study is restricted to the analytical application of chemiluminescence arising during the course of a chemical reaction, and bioluminescence a special type of chemiluminescence found in biological system in which a catalytic protein increase the efficiency of the chemiluminescence reaction. Light is produced when molecules reached to an excited state, and decayed to the ground state. We try to use luminous bacteria to create a novel biosensing system : Bioluminescent biosensing system in our laboratory. The study can be separated several parts : the construction of bioluminescent biosensing system, the confirm of the expression vector and host cell; the physiological condition test of the optimal host cell; the optimal test condition of the bioluminescent biosensing system; the actual test of the toxic compound. Our research construct expression vector (pUC-luxAB) containing luciferase gene, and transformed into E. coli BL21 cell. The optimal incubation condition was : TB broth, 27℃, 18hrs. The bioluminescent biosensing system test condition was : test temperature 35℃, using 100mL bacterial solution and 60mL injection substrate's volume (10% n-Decyl aldehyde was soluble in 9.5% ethanol) to produce the best bioluminescence. We also select toxic molecules such as formaldehyde and EPN to detect thesis toxicity in our experiment. According to experiment data, it shows there is a linear correlation between the inhibition rate of bioluminescence and concentration of toxic molecules : formaldehyde and EPN, when their concentration range from 100 to 700ppm, 6.5 to 9.5ppm respectively.