鲇鱼线粒体DNA的酶切图谱

STUDIES ON RESTRICTION ENZYME MAP OF MITOCHONDRIAL DNA FROM SILURUS ASOTUS

采用差速离心法及ＤＮａｓｅⅠ、ＲＮａｓｅ消化法制备并纯化了鲇（Ｓｉｌｕｒｕｓａｓｏｔｕｓ）肝脏线粒体ＤＮＡ（ｍｉｔｏｃｈｏｎｄｒｉａｌＤＮＡ，ｍｔＤＮＡ）。用８种限制性内切酶对ｍｔＤＮＡ进行了分析。ＢｇⅡ、ＥｃｏｒⅠ、ＰｓｔⅠ、ＢｇｌⅠ、ＢａｍＨⅠ、ＸｂａⅠ、ＨｉｎｄⅢ、ＸｈｏⅠ在鲇ｍｔＤＮＡ分子上分别具１、１、１、２、２、７、７和０个切点。ｍｔＤＮＡ分子量约１０．８４×１０￣６道尔顿，大小为１７．５４ｋｉｌｏｂａｓｅｐａｉｒｓ。根据单酶和双酶解片段的数目和分子量，建立了鲇ｍｔＤＮＡ的限制性酶切图谱。

The mitochondrial DNA (mtDNA) from liver of Silurus asotus had been isolated and purified by the method of density gradient centrifugation and DNase i, RNase digestion. The 8 kinds of restriction endonucleases, which were Xho Ⅰ, Bgl Ⅱ, EcoR Ⅰ,Pst Ⅰ, Bgl Ⅰ, BamH Ⅰ, Xba Ⅰ and Hind Ⅲ, had been used to digest the mtDNA. On the S. asotus mtDNA, the digested sites of these enzymes are 0, 1, 1, 1, 2, 2, 7 and 7,respectively. The molecular weight of S. asotus mtDNA is about 10. 84 ×106 and 17. 54 kilobase pairs. According to the number and the size of the fragment obtained by digested with single and double enzymes, the restriction enzyme map of mtDNA from S. asotus was constructed.