山羊-绵羊异质体细胞克隆胚中线粒体DNA的杂合性分析  

Mitochondrial DNA Heteroplasmic Analysis in Goat-sheep Heteroplasmic Somatic Nuclear Transfer Embryos

作　　者：杨丽[1] 马利兵[1] 华松[1] 曹俊伟[1] 张涌[1] 

机构地区：[1]西北农林科技大学生物工程研究所,杨凌712100

出　　处：《农业生物技术学报》2007年第4期628-632,共5页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2001AA213081)资助

摘　　要：采用PCR-限制性片段长度多态性(PCR-restriction fragment length polymorphism,PCR-RFLP)的方法分析以山羊(Capra hircus)胎儿成纤维细胞为核供体,以去核的绵羊(Ovisaries L.)MⅡ期卵母细胞为核受体构建的异质克隆胚中线粒体的来源及组成比例。结果表明,8-细胞之前的异质克隆胚中供受体mtDNA的比例变化不大,1-细胞、2-细胞、4-细胞和8-细胞异质克隆胚中核供体细胞线粒体DNA(mtDNA)占受体细胞mtDNA的比例分别为(2.5±0.8)%、(2.8±0.6)%、(1.9±0.4)%和(1.7±0.6)%,彼此之间差异不显著(P>0.05);但异质克隆囊胚中核供体细胞mtDNA的比例下降为(0.3±0.1)%,与前四者相比差异显著(P<0.05)。Goat（capra hircus）-sheep（Ovis aries L.） cloned embryos were constructed by fusing goat fetal fibroblasts （GFFs） into sheep oocytes, and then cultured in vitro to investigate the mitochondrial DNA （mtDNA） origin and proportion in reconstructed em bryos. Moreover, at each stage of 1- （immediately after fused）, 2-, 4-, 8-cell and blastocyst, the copy number of mtDNA from GFF and sheep oocyte were examined by using PCR-restriction fragment length polymorphism（PCR-RFLP）. The result showed that the propor- tions of nuclear donor to recipient mtDNA in l-cell,2-cell,4-cell and 8-cell reconstruction embryos were （2.5±0.8） %, （2.8±0.6） %, （1.9±0.4） % and （1.7±0.6） %, respectively, and the differrence was not significant （P 〉 0.05）. However, in heteroplasmic blastocysts, the proportion of nuclear donor mtDNA declined to （0.3±0.1）%, and the difference was significantly compared with that before 8-cell stage（P 〈 0.05）.

关 键 词：异种体细胞核移植 线粒体DNA 杂合性分析 山羊 绵羊 

分 类 号：S185[农业科学—农业基础科学]