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狼山鸡-AA肉鸡嵌合体的制作及其转基因鸡研究

DOI: 10.7685/j.issn.1000-2030.2007.02.017, PP. 83-88

Keywords: 胚盘细胞,嵌合体,转基因,狼山鸡

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Abstract:

试验Ⅰ以狼山鸡为供体,AA肉鸡为受体,通过赤道开窗法制作了狼山鸡-AA肉鸡的嵌合体。共操作了108枚鸡蛋,孵化成活19只,孵化成活率17.6%。获得表型嵌合体鸡胚22枚,其中15只孵化成活,嵌合体率为13.9%。对2只嵌合体公鸡解剖发现,有1只的睾丸发育异常。PCR扩增禽类W染色体特异性的重复序列发现,2只嵌合体公鸡的性腺都嵌合了供体异性的细胞。结果表明所采用的嵌合体制作方法制备转基因鸡是可行的。试验Ⅱ利用脂质体介导法,在体外将外源pEGFP质粒转入到鸡X期胚胎细胞中,培养8h开始有外源基因的表达,体外培养12h后获得了21.43%的转染效率。将转染后的鸡胚盘细胞移入到受体AA肉鸡的胚下腔,孵化6d后在8个鸡胚中检测到有外源基因的存在,阳性率为40%(8/20)。

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