This thesis introduced the principle and the evolution of polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), described its usage in monitoring the remediation process for oil and other chemical pollution as well as analyzing the nature microbial community, and finally made a comprehensive comment to its future development. PCR- DGGE was initially developed to study DNA mutations because it is a powerful tool to detect DNA base mutations in a PCR-amplified DNA fragment. Its accuracy can be as high as 100%. The principal of DGGE is based on the differences in DNA sequence that affect the melting points of each amplicons, and caused a decrease in the electrophoretic mobility of a partially melted DNA molecule in a polyacrylamide gel containing a linearly increasing gradient of DNA denaturants. PCR-DGGE can effectively detect the community structure of microorganisms in the environments, including the unculturable microorganisms. Nowadays, it has become one of the most frequently applied techniques to study the community structure of microorganisms. Microbiologists can use this technique to understand the microbial ecology, the shift of the microbial community structure during a bioremediation process, such as the oil pollution and other toxic chemical pollution. In addition, PCR-DGGE can also be used for the fermentation studies in food industry and agricultural industries. We can even identify unculturable microorganism by analyzing the DNA fragment sequences. This can help us to design a suitable medium to culture and isolate these ¡¥unculturable organisms¡¦. There are three directions for the further development of PCR-DGGE technique: 1.Improvement of material preparation processes; 2. development of new DGGE-related tools; and 3. combination of other technology with DGGE. The improvement of preparation processes can make DGGE more accuracy to analyze the community structure of microorganisms, improve the sensitivity of DGGE fingerprint detection, and help the DGGE normalization. The development of DGGE-related tools can overcome the limitations imposed on DGGE by co-migration and help us to have a better understanding of the meanings for a DGGE pattern. The combination of other technology with DGGE can help us to have a clear understanding of the microbial ecology, the shift of the microbial community structure, and the function of microorganisms.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0912106-104600 |
| Date | 12 September 2006 |
| Creators | Hsu, Po-an |
| Contributors | Jong-Kang Liu, none, Jimmy C. M. Kao |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0912106-104600 |
| Rights | off_campus_withheld, Copyright information available at source archive |
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