方正银鲫、白鲫与鲫线粒体DNA限制性内切酶酶切比较

COMPARATIVE STUDY ON RESTRICTION ENDONUCLEASE DIGESTION OF mtDNAs FOR CARASSIUS AURATUS GIBELIO,CARASSIUS AURATUS CUVIERI AND CARASSIUS AURATUS AURATUS

用 Bam HI、Eco RI、Dra I、Hinf I、Hind III、Hpa II、Msp I、Pst I、Sau 3AI、Sma I、Xba I、Xho I和 Sal I等十三种限制性内切酶对方正银鲫(Carassius aur-atus gibelio)、白鲫(Carassius auratus cuvieri)和鲫(Carassius auratus auratus)的线粒体 DNA(mt DNA)进行单酶酶切以及其中八种识别六碱基对序列的限制性内切酶的双酶酶切,经琼脂糖凝胶电泳后,分析且计算出各酶切片断大小,得出三种鲫鱼的 mt DNA分子大小:方正银鲫为 15990±90碱基对(bp);白鲫为 16600±130碱基对(bP);鲫为 15540±140碱基对(bp),并且分别建立了方正银鲫、白鲫和鲫 mt DNA由 Bam HI、Pst I、Eco RI 及 Xba I等四种限制性内切酶构建的酶切图谱。

The mitochondrial DNAs (mtDNAs) of Carassius auratus yibelio,Carassius auratus cuvieri and Carassius auratus auratus were digested by thirteenrestriction endonucleases, viz. Bam HI, Eco RI, Den Ⅰ,Hinf Ⅰ,Hind Ⅲ,Hpa Ⅱ,Msp Ⅰ,Pst Ⅰ, Sau 3AI, Sma Ⅰ, Xba Ⅰ, Xho Ⅰ and Sal Ⅰ. The double enzyme digestions wereconducted by using Bam HI, Eco RI, Hind Ⅲ, Sma Ⅰ, Xba Ⅰ, Sal Ⅰ and Pst Ⅰ. Thefragments were obtained and their molecular sizes were estimated by agarose gelelectrophoresis. The size of mtDNAs for C. auratus gibelio, C. auratus cuvieri and C.anratus auratus are 15990±90bp, 16600±130bp and 15540±140bp respectively. Theexperimental results demonstrated that the electrophoretic patterns of mtDNAsfor C. auratusgibelio, C. auratus cuvieri and C. auratus auratus obtained by eithersingle or double enzyme digestion were significantly different, and the patternsof C. auratus yibelio were obviously different among individuals by using Hpa Ⅱ.Besides, the restriction maps of mtDNAs from above species were constructed byusing Barn HI, Pst Ⅰ, Eco RI and Xba Ⅰ.