暗纹东方鲀mtDNA的分离纯化及其细胞色素b基因的分子克隆

Isolation and Purification of Mitochondrial DNA and Cytochrome b Gene Cloning in Obscure Puffer

利用改进的差速离心法和碱裂解法制备并分离纯化了暗纹东方mtDNA,测得分子大小为19.4Kb。用线粒体特异染色法跟踪和监测线粒体提取,结果表明,25000～35000g能获得90%以上的高得率。利用制备纯化的mtDNA,已首次克隆了暗纹东方细胞色素b基因(cytb)。

Mitochondrial DNA (mtDNA) was isolated and purified from obscure puffer Fugu obscurus by differential centrifugation and alkali degradation method. It was found that the mtDNA had molecular length of 19.4 kb. The procedure checked by Jenus Green B Staining Method step by step showed that more mitochondria (more than 90%) was obtained at centrifugation of 25000～35000 g. The cytochrome b (cytb) gene in the puffer was cloned for the first time.