碩士 / 國立臺灣大學 / 漁業科學研究所 / 85 / The green fluorescent protein(GFP) from Aequorea
victoria is capable of producing flourescence without an
exogenously added substrate. Here, we demonstrate that a cDNA
for GFP deriven by human CMV immediate early enhancer/promoter(
pGreen LANTERN)can confer flourescence upon live medaka embryos.
Threedifferent DNA concentration, 5ug/ml, 7.5ug/ml, 10ug/ml,
were injected into thegerminal vesicle of medaka oocytes, and
the GFP can be detected at 12 hr. afterartificial fertilization(
A.F.). To compare the survival rate at 24 hr. after A.F., all
embryos injected high DNA concentration(10ug/ml) were abnormal.
The green fluorescence of GFP in the embryos injected 7.5ug/ml
was stable and detectable at least 10 days after hatching out.
No green fluorescence in the embryos injected PBS. So, GFP cDNA
should be a good reporter gene to study thedynamics of gene
expression in medaka. The upstream regions, 3.5kb and
0.5kb, of rhodopsin gene from common carp were ligate into the
GFP expression vecter to study the dynamics of expression in the
transgenic medaka embryos. The expression of GFP in injected
upstream 3.5kb fragment embryos can be detected at 18 hr. after
A.F.. But, the green flourescence was transient disapper at 48
hr. after A.F.. At 72 hr., the GFP can be detected ,again, and
the expression of GFP only located at the eyes.But, the
expression of GFP in the embryos injected upstream 0.5kb
fragment wereweaker and non-tissue specific. The response
elements to control the tissue-specific expression should be
locate at the upstream 0.5~3.5kb region. The upstream
region 3kb of tilapia c-ski gene was ligate into GFP expression
vecter. And the expression of GFP can be detected at 12 hr. A.F.
inthe thansgenic medaka embryos. After the gastrula stage, the
GFP expression was only detecable at the yolk sac. Until
hatching out, the GFP was disappear.At 5th day after hatching
out, the yolk sac is disappear, and the GFP was detectable,
either. The function of tilapia c-ski gene maybe relate to the
development of yolk sac.
| Identifer | oai:union.ndltd.org:TW/085NTU00451008 |
| Date | January 1997 |
| Creators | Chung, Chih-Ching, 鍾至青 |
| Contributors | Tsai Huai-Jen, 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 | 70 |
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