鸡蛋开窗法对嵌合体鸡制备效率的影响  

Effect of Drilling a Window for Egg on the Efficiency of Chicken Chimeric Preparation

作　　者：王颖洁[1] 张文慧[1] 左其生[1] 李东[1] 张蕾[1] 汤贝贝[1] 连超[1] 肖天荣 张亚妮[1] 李碧春[1] 

机构地区：[1]扬州大学动物科学与技术学院江苏省动物繁育与分子设计重点实验室,江苏扬州225009

出　　处：《中国家禽》2015年第12期5-8,共4页China Poultry

基　　金：国家自然科学基金项目(31272429;31301959;31472087);高等学校博士学科点专项科研基金资助课题(20123250120009)

摘　　要：将受体种蛋分为3组,分别经换壳法、钝端开窗法和赤道面开窗后,比较3种开窗方法对孵化率的影响;分离培养鸡胚胎干细胞(ESCs),体外培养传代后用线性化的质粒p EGFP-N1转染鸡ESCs,比较显微注射时3种不同的处理方法对孵化率的影响;对获得的嵌合体鸡分别提取血液和组织DNA后进行PCR检测。结果表明:赤道面开窗法的孵化率显著高于换壳法和钝端开窗法(P<0.01);开窗后,显微注射经转染的ESCs组孵化率显著低于其他2组(P<0.01);PCR检测结果显示,有四只嵌合体鸡的部分器官表达了EGFP基因,其中两只鸡的性腺发生EGFP基因的嵌合。表明赤道面开窗后,对受体鸡胚下腔显微注射经转染的ESCs可以生产嵌合体鸡,产生嵌合体鸡的效率为4.17%。The recipient eggs was divided into three groups and the shedding method,drilling a window at the blunt end of egg and the lateral shell of egg was conducted, respectively, followed was the comparison of the hatchability among those three groups. Chicken embryonic stem cells （ESCs）were separated and cultured in vitro, then transfected with linearized plasmid pEGFP-N1. PCR was performed to detect the EGFP integration for the live chicken. The results showed that the hatchability of drilling a window at the lateral shell of egg was significantly higher than that of the other two group （P〈0.01）; The hatchability of microinjection with transfected ESCs was significantly lower than that of the other two group （P〈0.01）;PCR results showed four of the five live chicken expressed EGFP gene in some organs,but only two chimeric chicken expressed EGFP gene in the gonad. In all,the chimeric chicken could be produced by micro-injection ESCs in to chick embryo cavity after drilling a window at the lateral shell of egg,the efficiency was 4.17%.

关 键 词：开窗法 显微注射 嵌合体鸡 

分 类 号：S831.3[农业科学—畜牧学]