魚類基因轉殖及生長,發育,環境適應之研究(Ⅱ)─魚類之生長調控:SKI基因的調控及轉殖(Ⅱ)
Date Issued
1999-07-31
Date
1999-07-31
Author(s)
蔡懷楨
DOI
882311B002034B24
Abstract
Because green fluorescence protein (GFP) is a reportor gene used for in vivo
detection, we constructed a GFP cDNA driven by human cytomegalovirus immediate
early enhancer and promoter and microcinjected into medaka oocytes at various
concentrations. Results showed that GFP was detectable starting at embryonic stage 15;
the expression rates of 5μg/ml, %5μg/ml, and 10μg/ml groups were 50, 63 and 100%,
respectively. However, the suvival rates of these GFP embryos after stage 19 were 40, 50
and 0%, respectively, suggesting too high DNA concentration of microinjected DNA
caused abnormal development of embryos. The GFP was detectable until 10 days after
hatching without disturbing by endogenous green light for embryos microinjected with
7.5μg/ml.
Thus, we constructed a DNA fragment containing upstream 3 kb segment of tilapia
c-ski gene fused with GFP cDNA and microinjected into medaka embryos at 7.5μg/ml.
Resulets showed that GFP started to express at stage 15, but GFP was expressed
exclusively at yolk sac at gastrula stage 34 and last until hatching out. When yolk sac was
absorbed gradually the GFP disappeared gradually at the same time, and GFP was totally
abolished at 5 days after hatching. Results suggest that ski protein of lilapia may involve
in the development of yolk sac at early embryonic stag of fish.
Publisher
臺北市:國立臺灣大學漁業科學研究所
Type
journal article
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