碩士 / 國立臺灣大學 / 漁業科學研究所 / 86 / Sperm mediated gene transfer is a
rapid and convenient method to transfer foreign genes into organisms. A large
amount of samples could be treated in a short period of time. Also, the probl
ems we would encounter when transferring genes into eggs with tough egg shells
could be eliminated. In our study, we assessed the feasibility of transferrin
g foreign genes into Macrobrachium rosenbergii with this method. The DNA fragm
ent used in the experiments is the promoter of CMV (cytomegalovirus) immediate
early geneollowed by cDNA of GFP and SV40 polyadenylation signal in sequence.
The GFP gene is used as a reporter gene here. In the results of our experimen
ts. (1). The spermatophore could be obtained from male shrimp by using 7.5 vol
ts of electrical shock. No permanent injuries are observed after this treatmen
t. (2). Using ddH2O as the solvent in dissolving DNA samples would not affect
the fertilization and subsequent development of shrimp eggs. (3). When the for
eign DNA concentration exceeds 50ng/l, the eggs wouldot be nature hatched. T
he fertilization is successful as the concentration of DNA aqueous solution is
down to 28ng/l. But, the hatch rate is down to 42.5% (Hatch rate of the con
trol group is 82.5%). (4). We extract genomic DNA from 20 shrimps (at zoea 1 s
tage) which were treated with 28ng/l of DNA solution. After PCR amplificatio
n and southern blot analysis, we observed that a sample (5%) shows the same po
sitive signal with positive control. Their molecular weight are both 650bp. Th
erefore, the feasibily of transferring foreign DNA into Macrobrachium rosenber
gii is definite.
| Identifer | oai:union.ndltd.org:TW/086NTU00451010 |
| Date | January 1998 |
| Creators | li, si shen, 李思賢 |
| Contributors | Huai-Jen Tsai, 蔡懷楨, --- |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 1 |
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