博士 / 國立中興大學 / 動物科學系所 / 100 / In the present study, Both of novel family- and genus-specific DNA markers in Mugilidae fish and golden threadfin bream (Nemipterus virgatus). Genomic DNA was isolated from the blood of fish of 15 families and eighty random primers were used for random amplified polymorphic DNA (RAPD) fingerprinting. When the primer OPAV04 was employed, a novel specific PCR product was observed in the Mugilidae family. In addition, another novel specific PCR product was also observed in the Liza genus while using primer OPAV10. Sequencing analysis revealed that the novel family- and genus-specific DNA fragments were 856 bp and 418 bp, respectively, and no similar sequence was found in GenBank. Two primer sets were designed based on the family- and genus-specific sequences to confirm the RAPD results and the 569 bp and 186 bp predicted bands were successfully amplified by PCR. Intriguingly, Mugilidae family specific DNA markers were also effectively used for terrestrial and aquatic animal discrimination. A species-specific band amplified from Nemipterus virgatus using the OPAA11 random primer was selected for sequencing analysis, and a 535 bp sequence which had no similarity with any others in the nucleotide database was obtained. A primer set was designed based on the Nemipterus virgatus specific sequence to validate the RAPD-PCR results. A predicted 372 bp amplicon was significantly observed by PCR. Therefore, the novel family- and genus-specific DNA markers identified in this study can be used as an effective tool for rapid and accurate determination of which fish, and even for cross-species identification.
| Identifer | oai:union.ndltd.org:TW/100NCHU5289035 |
| Date | January 2012 |
| Creators | Shan-Hu Lai, 賴珊湖 |
| Contributors | Mu-Chiou Huang, 黃木秋 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 54 |
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