SSCP analysis of pig mitochondrial DNA D-loop region polymorphism |
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| Authors: | K Takeda A Onishi N Ishida K Kawakami M Komatsu S Inumaru |
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| Institution: | Department of Animal Breeding and Genetics, National Institute of Animal Industry, Norindanchi, Tsukuba, Ibaraki 305, Japan;Equine Research Institute, Japan Racing Association, Setagayaku, Tokyo 154, Japan;Institute of STAFF, Oazakamiyokoba, Tsukuba, Ibaraki 305, Japan;Biological Products Research Division, National Institute of Animal Health, 3-1-1 Kannondai, Tsukuba, Ibaraki 305, Japan |
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| Abstract: | The sequence polymorphism that occurs in the mitochondrial DNA (mtDNA) displacement (D)-loop region is useful as a cytoplasmic DNA marker. We cloned the mtDNA D-loop regions of five breeds of pig by polymerase chain reaction (PCR) and determined their sequences. The sequence diversities in D-loop regions among five breeds of pig were located in the starting area of heavy-strand replication. From these sequences, we designed primers for PCR-mediated single-strand conformation polymorphism (PCR-SSCP) analysis that amplified the most polymorphic 227 bp fragment of the D-loop region. The results of PCR-SSCP analysis clearly showed that four types of polymorphism (A to D) are found in Landrace (A), Large White (A, B), Duroc (A), Göttingen miniature pig (B) and Meishan (C, D). The same polymorphisms were also detected from each porcine embryo by this method. Our results show that PCR-SSCP analysis is useful in detecting polymorphisms in the D-loop region of pigs and pig embryos. |
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| Keywords: | D-loop region mitochondrial DNA PCR-SSCP analysis polymorphism porcine |
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