使用基于聚合酶链反应的方法检测牛线粒体DNA D环区域的核苷酸变异。

Detection of nucleotide variations in the D-loop region of bovine mitochondrial DNA using polymerase chain reaction-based methodologies.

作者信息

Eledath F M, Hines H C

机构信息

Department of Dairy Science, Ohio State University, Columbus 43210, USA.

出版信息

Anim Genet. 1996 Oct;27(5):333-6.

PMID:8930074

Abstract

Allele-specific polymerase chain reaction (ASPCR) and single-strand conformation polymorphism (SSCP) were used to characterize 16 Holstein maternal lines. ASPCR was used to detect polymorphic nucleotides at eight different positions in the displacement loop (D-loop) of bovine mitochondrial DNA (mtDNA): 16022, 16057, 16074, 16231, 16247, 106, 169 and 363. ASPCR analysis of the maternal lines showed variations at nucleotide positions 106, 169, and 363 of the mtDNA. Within-line variation was observed in five maternal lines for nucleotide 363. SSCP analysis of the mtDNA D-loop region revealed variations that classified the maternal lines into six different genotypes. Based upon the variations observed by ASPCR and SSCP analysis, the animals representing the 16 maternal lines could be assigned to 10 different genotypic groups. These procedures provide a rapid, simple, non-radioactive, and reliable method of detecting polymorphism in the D-loop region of bovine mtDNA.

摘要

采用等位基因特异性聚合酶链反应（ASPCR）和单链构象多态性（SSCP）对16个荷斯坦母系进行特征分析。ASPCR用于检测牛线粒体DNA（mtDNA）置换环（D-loop）中八个不同位置的多态性核苷酸：16022、16057、16074、16231、16247、106、169和363。对母系的ASPCR分析显示，mtDNA的核苷酸位置106、169和363存在变异。在五个母系中观察到核苷酸363的系内变异。mtDNA D-loop区域的SSCP分析揭示了变异，这些变异将母系分为六种不同的基因型。基于ASPCR和SSCP分析观察到的变异，代表16个母系的动物可分为10个不同的基因型组。这些方法提供了一种快速、简单、非放射性且可靠的检测牛mtDNA D-loop区域多态性的方法。