武昌鱼肝线粒体DNA限制性内切酶酶解图谱与12S rRNA基因的初步定位

RESTRICTION MAP OF MITOCHONDRIAL DNA FROM MEGALOBRAMA AMBLYCEPHALA LIVER AND PRELIMINARY LOCALIZATION OF 12S rRNA GENE OF THE mtDNA

武昌鱼肝线粒体(mt)DNA经六种限制性内切酶BamHI,BgⅢ,BgⅡ,EcoRI,HindⅡHpall单酶完全酶解分別得到2,2,3,3,3和7个片段。用琼脂糖凝胶电泳测得各个酶解片段的长度和分子量,经计算该mtDNA长约16.6kb,分子量10.2×10~6道尔顿(dalton)。用七对限制酶双酶全酶解,构建出五种限制性内切酶图谱。以酵母线粒体15SrRNA基因为探针对武昌鱼肝mtDNA中的12SrRNA基因进行初步定位。

Wuchang Fish (M. amblycephala) liver mtDNA was cleaved by the restriction endonuclease. Upon complete digestion numbers of restriction fragments of BamHI, BglⅡ, BglⅠ, EcorⅠ, HindⅢ and HpaⅡ were determined to be 2,2,3,3,3 and 7 respectively. Molecular weight of the mtDNA was found to be 10×10~6 dalton or 16.6 kb. A restriction Map of five restriction endonucleases was constructed. 12S rRNA gene of the mtDNA was preliminarily localized by molecular hybridization using 15S rRNA gene from yeast mtDNA as the probe.