Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305 806, Republic of Korea.
Biochem Biophys Res Commun. 2012 Aug 10;424(4):765-70. doi: 10.1016/j.bbrc.2012.07.031. Epub 2012 Jul 15.
Somatic cell nuclear transfer (SCNT) has been established for the transmission of specific nuclear DNA. However, the fate of donor mitochondrial DNA (mtDNA) remains unclear. Here, we examined the fate of donor mtDNA in recloned pigs through third generations. Fibroblasts of recloned pigs were obtained from offspring of each generation produced by fusion of cultured fibroblasts from a Minnesota miniature pig (MMP) into enucleated oocytes of a Landrace pig. The D-loop regions from the mtDNA of donor and recipient differ at nucleotide sequence positions 16050 (A→T), 16062 (T→C), and 16135 (G→A). In order to determine the fate of donor mtDNA in recloned pigs, we analyzed the D-loop region of the donor's mtDNA by allele-specific PCR (AS-PCR) and real-time PCR. Donor mtDNA was successfully detected in all recloned offspring (F1, F2, and F3). These results indicate that heteroplasmy that originate from donor and recipient mtDNA is maintained in recloned pigs, resulting from SCNT, unlike natural reproduction.
体细胞核移植（SCNT）已被用于特定核 DNA 的传递。然而，供体线粒体 DNA（mtDNA）的命运尚不清楚。在这里，我们通过第三代研究了重编程猪中供体 mtDNA 的命运。重编程猪的成纤维细胞是通过将明尼苏达州微型猪（MMP）的培养成纤维细胞融合到长白猪去核卵母细胞中产生的每一代后代的成纤维细胞获得的。供体和受体 mtDNA 的 D 环区域在核苷酸序列位置 16050（A→T）、16062（T→C）和 16135（G→A）处存在差异。为了确定重编程猪中供体 mtDNA 的命运，我们通过等位基因特异性 PCR（AS-PCR）和实时 PCR 分析了供体 mtDNA 的 D 环区域。供体 mtDNA 在所有重编程后代（F1、F2 和 F3）中均成功检测到。这些结果表明，与自然繁殖不同，源自供体和受体 mtDNA 的异质体通过 SCNT 得以在重编程猪中维持。
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