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Improvement of the Fish Cold Acclimation
|Contributors: ||NTOU:Department of Aquaculture|
Tilapia;Carp;Creatine Kinase;Gene Transfer
|Issue Date: ||2011-06-28T07:45:24Z
|Abstract: ||摘要:熱休克蛋白Hsc70啟動子含有多個重要低溫表現相關調控區間。以此啟動子進行斑馬魚活體表現分析，可發現於16 ℃低溫下具有提升三倍表現量之能力，並於14 ℃低溫下仍保有表現能力。與病毒持續型啟動子CMV比較，低溫下如表現量相同則常溫下表現量約降為CMV表現量的1/6，可望大幅緩解常溫下持續大量表現對於能量異常消耗與正常生理機能之干擾。 今年將取得誘發抗寒機制所需之啟動子及肌肉特異性啟動子，減少多餘不必要的調控區段對於基因表現造成負面之影響，所選殖之啟動子後接螢光基因與抗寒相關酵素基因另可望提高轉殖基因的表現成功率與基因轉殖魚的存活率，並建立基因轉殖吳郭魚。|
Abstract:Heat shock proteins (Hsps) are well-known stress-inducible chaperons. It had been reported can be induced by cold shock in many species from plant to vertebrate. To understand the regulation of cold-induced gene expression, we identified the expression of zebrafish hsc70 by semi-quantitative RT-PCR, isolated the 5'-flanking sequence of zebrafish hsc70 gene and used a green fluorescent protein (GFP) for in vivo assay the promoter activity. In basal expression pattern, cold shock treatment result in an 10-folds enhanced induction of zebrafish hsc70 gene. In in vivo promoter assay, transient zebrafish shows an 2-folds enhanced induction while the constitutive CMV promoter reduced its expression level to 1/3 under cold shock treatment. We will identify and determine the muscle-specific promoter and get rid of the unnecessary region. The muscle-specific promoter is followed by green fluorescence protein or creatine kinase. Transgenic tilapia with creatine kinase will be produced and improved the fish in cold acclimated.
|Appears in Collections:||[水產養殖學系] 研究計畫|
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