碩士 / 國立海洋大學 / 水產養殖學系 / 88 / Since 1980, technique of transgenic was developing and the successful examples of transgenic animal were more than before. Transgenic animal such as: mouse, Dorsophila and C. elegans were had been succeeding.
In recent years, we use fish as a model animal for transgenic and because of their short life cycle, large number spawning and easy to observe their transparent fertilized eggs, especially zebrafish. We found that transgene in fish was unstable and will be loosed easily. And we track the expression and retention of transgene in fish. In the results, transgene could be found after fertilized 6 days but can't see that protein level have different expression between experiment group and control. We thought that might be the transgene do not integrate into the genome DNA and also can explain transgene why transgene unstable and eliminate day by day.
For this reason, we use Rec A protein to help transgene recombination and integrate into genome DNA. Rec A protein is exists in eukaryotes and prokaryotes. The main function of Rec A protein is DNA repair and help DNA recombination in SOS system. In mammal, there have succeeded to introduce homologous DNA into fertilized eggs with DNA recombination in mouse and sheep.
In our study, we use Rec A protein to help homologous DNA recombination and compare homologous DNA with no Rec A protein exist which one can help DNA recombinant or both can help it happen.
As the result, in the case of zebrafish that can’t help homologous DNA to recombinant with any Rec A protein. And we can find the group with Rec A protein can help our homologous DNA, which use electroporation into fertilized eggs happen recombination. The result can see under microscope, we found the red and green fluorescent is on the same site. Beside, we found the recombination was happened at the specific site and our tranegene was insert into the specific site, this is the most important discovery of transgenic research.
With this result, we can study gene expression and influence on different site of genomic DNA further.
| Identifer | oai:union.ndltd.org:TW/088NTOU0086007 |
| Date | January 2000 |
| Creators | Chi I Lee, 李芷懿 |
| Contributors | Jau-Der Chen, 陳昭德 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 94 |
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