Establishment of molecular approaches to detect the nitrous oxide reductase containing microbes in aquatic environments

• Main Author: 劉宏達
• Other Authors: 應靜雯
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/24801929902403420079

碩士 === 東吳大學 === 微生物學系 === 91 === The N2O is involved in global warming and the destruction of the ozone layer. Denitrification is also used in waste treatment facilities to remove excess combined nitrogen. Denitrification is a process that involves all the major physiological groups. It has been difficult to detect the numbers of these microbes via classic approach. The conversion of N2O to N2 is the last step of a complete denitrification pathway. Based on the nucleic acid sequence of nosZ gene reported, several primers specific for this gene were designed and evaluated. Using the primers reported previously, a single unique DNA fragment with a molecular weight of 700bp can be amplified in nosZ containing bacteria. The sensitivity and detection limit of this system is about 103 denitrifers. We have evaluated the application of PCR-ELISA on the quantification of the nitrous oxide reductase denitrifers in the aquatic samples. By the PCR-ELISA system , a linear range of 5x103~105 denitrifers can be detected in the aquatic environments samples.