白血病细胞 MtDNA RFLP 分析与D-环片段的制备

Analysis of RFLP and Preparation of D Loop on MtDNA in Human Leukemic Leucocytes

从正常人胎盘组织和白血病外周血细胞分别提取线粒体ＤＮＡ，选用ＢａｍＨＩ／ＳｐｈⅠ双酶切线粒体ＤＮＡ，琼脂糖凝胶电泳分离，分析酶切产物，以λ－ＤＮＡ－ＨｉｎｄⅢ为标准分子量对照。其ＲＦＬＰ结果表明，白血病人外周血线粒体ＤＮＡ的酶切产物为３个片段，而正常人胎盘组织线粒体ＤＮＡ的酶切产物为两个片段，提示白血病人的线粒体ＤＮＡ发生了结构改变，同时用低熔点琼脂糖凝胶回收含Ｄ－环的线粒体ＤＮＡ片段。本研究为研究白血病与线粒体ＤＮＡ的相关性提供了实验依据。

Mitochondrial DNA(Mt DNA) in normal human placenta and human leukemic peripheral blood cells has been extracted.The MtDNA was digested with restriction endonuclease BamHI and SphI,the digested samples were electrophoresed on agarose gels and DNA HindⅢ as standard.The results of RFLP showed that digested MtDNA in human leukemic peripheral cells has three fragments,digested MtDNA in normal human placenta has two fragments.It indicated that the structures of MtDNA in human leukemic leucocytes have been changed.D loop of MtDNA was obtained by low melting temperature agarose electrophoresis.Our studies will provide evidences for relativity of leukemic and MtDNA.