RAPD Fingerprinting for Species Identification of Animals

• Main Authors: Chia-Yu Lin, 林佳瑜
• Other Authors: Mu-Chiou Huang
• Format: Others
• Language: zh-TW
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/29104389683697974491

碩士 === 國立中興大學 === 畜產學系 === 92 === 一、Species Identification of Fowls Abstract The studies were based on the RAPD fingerprinting for the species identification of fowls. There were 12 kinds of fowls being identified in this study. The genomic DNA samples of animals were employed as the templates to amplified with random primers by RAPD-PCR for fingerprinting. The species-specific fragments were isolated from gel and ligated into vector for nucleotide sequencing. Designed the species-specific primer on the basis of sequence analysis and performed PCR reaction to identify the species of animals. The results showed that the RAPD fingerprinting pattern of fowls revealed a significant diversity, but the RAPD fingerprinting band in the same species of fowls (such as Leghorn, Shaver and Native chicken) showed a highly similarity. Some species-specific fragments were represented in the RAPD fingerprints of ostrich, emu, turkey, Muscovy duck, goose and pigeon, which were amplified with OPAV-05, OPAA-10, OPAO-10, OPAV-13, OPAO-07 and OPAV-19 primers. Primers of OstrAV5SpeF3 and OstrAV5SpeR2 were designed according to the cloned species-specific sequence of ostrich, which were employed for PCR with the template DNA of fowls, a 295 bp species-specific band was found in ostrich only. With the design of the EmuAA10SpeF1 and EmuAA10SpeR1 species-specific primers from the species-specific sequence of emu, which were employed for PCR with the template DNA of fowls, a 501 bp species-specific band was found in emu only. With the design of the LegAA10SpeF1 and the LegAA10SpeR1 species-specific primers from the species-specific sequence of Leghorns, which were employed for PCR with the template DNA of fowls, a 358 bp species-specific band were found in Leghorns, Shavers, and Native chicken, but band wouldn’t be generated within turkeys and other fowls, so the primer had the chicken species-specific diversity to be distinguished between chickens and turkeys. Primers of TurkAV19SpeF1 and TurkAV19SpeR1 were designed according to the cloned species-specific sequence of turkey, which were employed for PCR with the template DNA of fowls, a 203 bp species-specific band was found in turkey only. Primers of MuscAO10SpeF3 and MuscAO10SpeR3 were designed according to the cloned species-specific sequence of Muscovy duck, which were employed for PCR with the template DNA of fowls, a 785 bp species-specific band was found in Muscovy duck only. Primers of WRomAO7SpeF1 and WRomAO7SpeR1 were designed according to the cloned species-specific sequence of White roman goose, which were employed for PCR with the template DNA of fowls, a 219 bp species-specific band were found in White roman goose and Chinese goose only. Primers of PigeAV13SpeF1 and PigeAV13SpeR1 were designed according to the cloned species-specific sequence of pigeon, which were employed for PCR with the template DNA of fowls, a 150 bp species-specific band was found in pigeon only. 二、Species Identification of Farm Animals Abstract The studies were based on the RAPD fingerprinting for the species identification of farm animals. There were 12 kinds of farm animals in this study. The genomic DNA samples of animals were employed as the templates to amplified with random primers by RAPD-PCR for fingerprinting. The species-specific fragments were isolated from gel and ligated into vector for nucleotide sequencing. Next, designed a species-specific primer on the base of DNA sequence analysis to proceed PCR reaction in order to identify the animal breed. The band was similar within the same species (such as Yellow cattle, Holstein cattle and Angus cattle) form the RAPD fingerprinting pattern of farm animals, even when the fingerprint pattern of cattle and sheep would generate a similar band, which was probably because these two species were close to each others’ ruminate relation. The RAPD fingerprint among pigs, cattle, and sheep had a major significant diversity. Some species-specific fragments were represented in the RAPD fingerprints of Yellow cattle, Yorkshire pig and Native goat, which were amplified with OPAD-12, OPAO-19 and OPAE-10 primers. Primers of YcatAD12SpeF1 and YcatAD12SpeR1 were designed according to the cloned species-specific sequence of Yellow cattle, which were employed for PCR with the template DNA of farm animals, a 619 bp species-specific band was found in cattle (Yellow cattle, Water buffalo, Holstein cattle, and Angus cattle) only. Primers of YorkAO19SpeF1 and YorkAO19SpeR1 were designed according to the cloned species-specific sequence of Yorkshire pig, which were employed for PCR with the template DNA of farm animals, a 393 bp species-specific band was found in pig (Duroc pig, Landrace pig, Yorkshire pig and Hampshire pig) only. Primers of NaGotAE10SpeF2 and NaGotAE10SpeR2 were designed according to the cloned species-specific sequence of Native goat, which were employed for PCR with the template DNA of farm animals, a 168 bp species-specific band were found in sheep and goat (Dorset sheep, Native goat, Nubian goat and Milk goat) only. In other word, those species-specific primer designs above are applied to species identification of different kinds of animals such as cattle, pigs, sheep and goat.