摘要
本文报道配合使用差速离心和DNaseI核酸酶处理等步骤,从草鱼和鲤鱼新鲜肝组织分离线粒体DNA(mtDNA)的实验方法。这种方法经济简便,纯化的mtDNA产率多,纯度高,经限制性内切酶消化后进行琼脂糖凝胶电泳分离,可以得到清晰的DNA片段谱带,并可直接用于构建酶切图谱和线粒体基因的分子克隆。用这样的mtDNA,我们已克隆了草鱼和鲤鱼的细胞色素氧化酶亚基I基因(COI基因)。
Here we report a procedure for the isolaion and purification of mitochondrial DNA (mtDNA) from fresh livers of grass carp (Ctenopharyngodon idellus) and carp (Cyprinus carpio L.) by differencial centrifugation and DNaseI treatment. The procedure is economical and convenient. The mtDNA obtained by this procedure showed high purity and high yield. The grass carp mtDNA was digested by several restriction enzyme and clear bands were shown on agarose gel electrophoresis. The purified mtDNA can be used directly for mapping of mtDNA and molecular cloning of mitochondrial genes. We had successfully cloned mitochondrial cytochrome oxidase subunit I (COI) gene from grass carp and carp.
出处
《动物学报》
SCIE
CAS
CSCD
1991年第4期375-382,共8页
ACTA ZOOLOGICA SINICA
关键词
草鱼
鲤鱼
线粒体
脱氧核糖核酸
Mitochondrial DNA, Restriction mapping, COI gene cloning.
