Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/25189
標題: 以體細胞核轉置技術產製基因轉殖山羊
Production of Transgenic Goats by Somatic Cells Nuclear Transfer
作者: 曲鳳翔
Chu, Feng-Shiang
關鍵字: nuclear transfer;核轉置;goat;transgenic;山羊;基因轉殖
出版社: 動物科學系所
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摘要: 
本研究旨在評估應用體細胞核轉置(somatic cell nuclear transfer技術產製轉基因複製(transgenic-cloned)山羊之可行性,探討其中影響轉基因核轉置山羊胚生產系統產製效率之卵母細胞來源、體外培養環境、供核細胞繼代次數等因子。除了生產帶有人類血清白蛋白(human serum albumin, hSA)之基因轉殖山羊之外,並能建立較穩定之轉基因核轉置山羊生產模式。若單純以建立特殊基因之生物反應器為目的,則乳山羊有多胎、體型小、乳量高、世代間距短、巨胎症與難產少及廢棄物易清理等多項優點。但乳山羊之族群數量少,不易取得試驗所需之大量卵母細胞。本試驗以人類血清白蛋白外源基因pGB562/hSA為標的基因,轉染優質山羊之體細胞株,並利用基因轉殖動物複製技術平台,以屠宰場收集並經體外成熟培養之羊卵母細胞及山羊體內收集之卵母細胞做為材料,產製具組織特異性表現外源基因蛋白的基因轉殖複製山羊。於屠宰場收集之卵母細胞經體外成熟培養,成熟率達63.7%,以其進行核轉置胚操作,共移置273個核轉置胚於18頭代孕母羊,其中一頭懷孕分娩,經基因檢測確定帶有HSA外源基因,產仔率5.6%。此外利用外科手術收集山羊體內成熟卵母細胞進行核轉置之結果,成熟率達77.15%,共移置417個核轉置胚於29頭代孕母羊,其中二頭懷孕分娩,產仔率6.8%。

The purpose of this study was to investigate the possibility of producing transgenic goat by somatic cell nuclear transfer technology. The influence of the source of oocyte, in vitro culture environment, and the number of passengers of donor cells on efficiency of production of transgenic nuclear transfer cloned goat were investigated. In addition, to produce the transgenic cloned goat carrying human serum albumin protein in milk the current research is to establish stable production system of transgenic cloned goat. If purpose of a study is only to consider establishing specific gene carried in animals, goat is a candidate animal which has many advantages, such as polyembryony, lighter body weight, higher milk production, shorter generations interval, lower chance of giant fetal disease and dystocia, and easier clean disposal. However, the population of goat is low, it is not easy to collect large amount of oocytes. The purpose of current study is to produce reconstruct transgenic cloned goat carrying specific gene such as pGB562/hAS of human serum albumin by using goat primary ear fibroblasts as nuclear donor cells transfected with pGB562/hSA and mature nucleated oocytes as receipt are collected from slaughter house. The in vitro maturation rate of goat oocytes was 63.7%. Total 273 nuclear transfer embryos were transferred into 18 surrogate mother goats. One goat had given birth and birth rate was 5.6%. Exogenous hSA gene was detected in this goat. In contrast to in vitro culture system, the mature oocytes were collected directly from ovary by surgery as in vivo system for nuclear transfer study. The maturation rate was 77.15%, and sum total of 417 nuclear transfer embryos were transferred to 29 surrogate mothers., then two goats had given birth, the birth rate was 6.8%.
URI: http://hdl.handle.net/11455/25189
其他識別: U0005-1808200911341300
Appears in Collections:動物科學系

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