洞庭青鲫与其他六个鲫鱼品系线粒体DNA控制区的比较分析
COMPARATIVE ANALYSIS OF THE MITOCHONDRIAL DNA CONTROL REGION IN DONGTINGKING CRUCIAN CARP AND OTHER SIX STRAINS OF CARASSIUS AURATUS
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摘要: 运用PCR扩增、克隆、测序等技术,在获得洞庭青鲫和彭泽鲫线粒体DNA控制区全序列的基础上,对洞庭青鲫、彭泽鲫、普通鲫、红鲫、白鲫、A系和D系银鲫等7个鲫鱼品系线粒体DNA控制区的碱基组成、变异情况、序列结构和系统进化进行了比较分析。结果表明,7个鲫鱼品系线粒体DNA控制区碱基的平均组成为A:(32.7±0.16)%,C:(20.4±0.37)%,G:(14.1±0.08)%,T:(32.8±0.36)%,序列分歧率为0—5.7%,其中红鲫和白鲫的分歧率最大(5.7%),彭泽鲫、A系和D系银鲫之间最小(0)。洞庭青鲫与白鲫的分歧率为5.6%,与彭泽鲫、A系和D系银鲫为2.2%,与红鲫和普通鲫分别为0.8%和0.7%。对比哺乳动物线粒体DNA控制区结构,并参照其他鱼类序列,将7个鲫鱼品系的控制区分为终止序列区、中央保守区和保守序列区三个区域。同时识别了7个鲫鱼品系中一系列保守序列,并给出了它们的一般形式。序列差异和系统进化分析表明,在7个鲫鱼品系中,洞庭青鲫与普通鲫和红鲫的亲缘关系最近,与彭泽鲫、A系和D系银鲫亲缘关系次之,与白鲫的亲缘关系最远,而彭泽鲫、A系和D系银鲫三者在线粒体DNA控制区的相似性和分歧率上则表现为是同一个鲫鱼品系。Abstract: In this study, the complete fragment of the mitochondrial DNA control region were amplified using the genomic DNA of Dongtingking crucian carp (Carassius auratus Dongtingking variety) and Pengze crucian carp (Carassius auratus of Pengze). The PCR products were ligated and sequenced to the plasmid of pMD18-T Vector by the method of direct cloning, and then, the base composition, variation, sequence structure and phylogeny of the mitochondrial DNA control region were compared and analyzed among seven strains of Carassius auratus (Dongtingking crucian carp, Pengze crucian carp, crucian carp, Red crucian carp, Japanese crucian carp, Silver crucian carp clone A and D). In the seven strains of Carassius auratus, the percentages of average base compositions of the mitochondrial DNA control region were A (32.7±0.16)%, C (20.4±0.37)%, G (14.1±0.08)% and T (32.8±0.36)%, respectively, the sequences percent divergences were between 0—5.7%, the highest percent divergence (5.7%) was between Red crucian carp and Japanese crucian carp, and the lowest percent divergence (0) was among Pengze crucian carp, Silver crucian carp clone A and D. The percent divergence between Dongtingking crucian carp and Japanese crucian carp was 5.6%, which was 2.2% among Dongtingking crucian carp and Pengze crucian carp, Silver crucian carp clone A and D, 0.8% between Dongtingking crucian carp and Red crucian carp, and 0.7% between Dongtingking crucian carp and crucian carp. Compared with the mitochondrial DNA control region in mammalian and other fishes, the extended terminal associated sequences (ETAS), central conserved sequence blocks (CSB-F, CSB-E, CSB-D), conserved sequence blocks (CSB-1, CSB-2, CSB-3) and putative promoters of the mitochondrial DNA control region in the seven strains of Carassius auratus were observed. Analysis of the sequences percent divergences and phylogeny indicated that the Dongtingking crucian carp had a extremely close relationship with crucian carp and Red crucian carp, taking the second place with Pengze crucian carp, Silver crucian carp clone A and D, and had a farthest relationship with Japanese crucian carp. Moreover, the consistent similarity and percent divergence of the mitochondrial DNA control region among Pengze crucian carp, Silver crucian carp clone A and D demonstrated that they were the same strain of Carassius auratus.
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