外源基因导入鸡胚的两种显微注射方法的比较  

Comparison of Two Methods of Introducing Foreign DNA into The Chicken Embryo

作　　者：孙研研[1] 郑江霞[1] 徐桂云[1] 曲鲁江[1] 杨宁[1] 

机构地区：[1]中国农业大学动物科学技术学院,北京100193

出　　处：《农业生物技术学报》2010年第5期938-943,共6页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2006AA10A121;No.2008AA101009);国家重大基础研究发展规划(973)项目(No.2006CB102102)共同资助

摘　　要：胚盘下腔显微注射和卵黄囊血管显微注射是向鸡胚中导入基因等外源物质的常用方法。本研究从实验操作难度、外源基因导入效果、对鸡胚存活率的影响等方面比较了这两种方法的优缺点。实验结果表明,胚盘下腔显微注射操作较简单,在鸡胚发育的不同日龄直至出雏后都可以检测到非整合的外源基因的存在,经胚盘下腔显微注射的鸡胚的存活率为9.5%;卵黄囊血管显微注射操作难度大,在不同日龄及出雏鸡只中也可以检测到外源基因的存在,外源质粒仍以游离状态存在,并未与宿主基因组发生整合,经卵黄囊血管注射的鸡胚的存活率为35.7%。两者各有利弊,研究中应根据实验目的、导入外源物质的作用时间、样本量的多少选择适当的注射方法。There are two major methods to introduce exogenous substances like foreign DNA into the chicken embryo.One is microinjection of the foreign DNA into the subgerminal cavity of fresh fertilized eggs before incubation.The other is microinjection of the foreign DNA into the yolk sac vein vessel after the fertilized egg incubated for 72 h.Workload,technical difficulty,effectiveness,and the hatchability of recipient chickens were listed as criteria to evaluate the two methods.The results indicated that the former way was easier to conduct but with a lower hatchability of 9.5%.The foreign DNA could be found in the recipient embryos of different ages and the chicks after hatching;The latter way was relatively difficult but with a higher hatchability of 35.7%.The foreign DNA could be found in the recipient embryos of different ages and the chicks after hatching,also without integration.Which way to be selected depends on the aim of the experiment,action time of the exogenous substances and the sample size.

关 键 词：鸡胚 胚盘 血管 显微注射 质粒 

分 类 号：S961.6[农业科学—水产养殖]