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Introduction of Green Fluorescent Protein and Lac-Z Gene into Zebrafish (Danio Rerio) Egg by Microinjection and Particle Gun

顯微注射與粒子槍基因轉殖法轉殖綠螢光蛋白與半乳醣甘酶于斑馬魚之研究

摘要


從水母中選殖出之綠螢光蛋白標的與其他生物分析標的具有與衆不同之性質,其具有潛在研究性質並可運用於魚類發生生物學,基因轉殖研究與水產養殖上。此研究利用顯微注射與粒子槍基因轉殖法轉殖pCMV-β質體進入斑馬魚授精卵,經染色分析得到80%與6%之表現率。在顯微注射綠螢光蛋白進入斑馬魚授精卵後具有80%表現率。顯微注射綠螢光蛋白(pEGFP-c1 vector)五小時後,可以看見綠螢光表現。綠螢光蛋白基因轉殖之斑馬魚可以在肌肉骨骼纖維,心臟,血球,骨髓,眼之活體上觀察到基因表現情形,並持續在42天大之斑馬魚肌肉上看到螢光蛋白之表現。

並列摘要


Green fluorescent protein (GFP) from Aequorea victoria is a novel marker compared with other bioassay markers that has potential use in the study of teleost development biology, transgenic research, and aquaculture. Microinjection and particle gun were used to transfer pCMV-β into fertilized zebra fish eggs. After staining analysis, the data showed80% and 6% expression rates, respectively. After microinjection of the pEGFP-C1 vector in to fertilized zebra fish eggs, the expression rate was 80%.GFP was visible 5 h after micro injection of the pEGFP-C1 vector. Following advancement of the development stage, the signal was visible in muscle skeletal fibers, heart, and corpuscles of living embryos. Transgenic fish continued to express high levels of GFP in bone marrow, eyes, and muscle in 42-d-old zebra fish as revealed by histological sections data.

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