Early Sex Identification on Holstein Embryo by Polymerase Chain Raction/Oligonucleotide Ligation Assay
| Autor: | CHIH-HAO KAO, 高志豪 |
|---|---|
| Rok vydání: | 2001 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 89 Advances in biotechnology in animal reproduction have freed livestock managers from many traditional breeding tasks, recently. These are major in embryo transfer techniques, in vitro fertilization (IVF), embryo freezing, embryo culture, and embryo sexing. Combined use of these techniques has greatly promoted efficiency in animal husbandry. Among them, sex identification of pre-implantation mammalian embryos played an important role in livestock management and breeding. It is potentially of great commercial importance to animal agriculture to control of the sex ration of domestic species. In order to achieve this goal, it must to develop a method of sex determination that would theoretically offer improved sensitivity and specificity. In the present reporter, we use a completely new strategy for sex determination of pre-implantation bovine embryo-Polymerase Chain Reaction/Oligonucleotide Ligation Assay (PCR/OLA). According to this method, we designed two adjacent oligonucleotides (20-mers) by the side of known single nucleotide polymorphisms (SNPs) on the highly conserved region of X-Y homologous gene-ZFX/ZFY (Zinc finger protein on chromosome X or Y). The two nucleotides are hybridized to target DNA and, if there are perfect complementarily, the enzyme DNA ligase covalently joins the 5’ Ligation probe and the 3’ Joining probe. Apply the principle of immunoassaying, it can be distinguished all variation from color presented in the wells by naked eye. Then, it can be used to identify sex of single blastomere from morula or early blastocyte of bovine embryo in proportion to 6-7 days after fertilization. Hoping increasing accuracy ratios of identification and decreasing injure of embryo from freezing. It not only be used to apply this system for sexing of bovine pre-implantation embryo but also can be used to others domestic animals. It also can be applied to detect inherited diseases or econamic trait. Finally, it expect high throughput, accuracy and fast identify sex of embryos by develop this analysis method for promoted efficiency in animal husbandry. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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