摘要
以太行山羊为研究对象,探索精子作为外源基因载体建立转FecB目的基因山羊的可行性,以及脂质体对转染效率的影响。采用共培养与阳离子脂质体介导2种方法转染精子,用原位杂交等技术检测FecB目的基因片段在精子中的存在及存在部位,并对2种方法进行分析。结果表明:太行山羊的精子具有主动捕获外源基因的能力,共培养法精子阳性率为18.3%,脂质体介导精子阳性率为38.7%,脂质体的存在显著提高了精子的转染效率。太行山羊的精子能够被脂质体包被的FecB目的基因片段转染,为精子载体法制备FecB转基因太行山羊提供依据。
To explore the feasibility of spermatozoa of Taihang goat as a vector of foreign genes to generate transgenic goats,and assess the effect of liposome during the spermatozoa transfection.Transfecting the spermatozoa by co-culture and liposome mediated methods with the FecB gene fragment.And two group samples were detected by PCR and FISH,and analyzed by statistics.The result showed that the spermatozoa could take exotic DNA voluntarily.The sperm positive rate of the total culture group was 18.3%;the sperm positive rate of liposome group was 38.7%.The transfection ratio of liposome mediated group was higher than co-culture group（P〈0.01）.The spermatozoa of Taihang goat could be transfected with the FecB gene fragment mediated by liposome and provided the basis of transgenic tai hang goat by SMGT.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2011年第1期97-100,110,共5页
Journal of Hebei Agricultural University
基金
国家科技部重大专项
优质转基因羊新品种培育(2008ZX08008-003)
