Inheritance of mitochondrial DNA in serially recloned pigs by somatic cell nuclear transfer (SCNT).
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.
Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305 806, Republic of Korea.
Minhwa
M
Do
Won-Gu
WG
Jang
Jeong Hee
JH
Hwang
Hoon
H
Jang
Eun-Jung
EJ
Kim
Eun-Jeong
EJ
Jeong
Hosup
H
Shim
Sung Soo
SS
Hwang
Keon Bong
KB
Oh
Sung June
SJ
Byun
Jin-Hoi
JH
Kim
Jeong Woong
JW
Lee
D000818:Animals; D001483:Base Sequence; D019976:Cloning, Organism; D004272:DNA, Mitochondrial / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism; D008969:Molecular Sequence Data; D053652:Nuclear Transfer Techniques; D009690:Nucleic Acid Conformation; D009865:Oocytes / Q000378:metabolism; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics*
D004272:DNA, Mitochondrial