258 CLASSIFICATION AND IDENTIFICATION OF THE CLONED HANWOO CALVES DERIVED FROM SOMATIC CELL NUCLEAR TRANSFER USING MS GENOTYPES AND mtDNA POLYMORPHISMS

Abstract

The calves produced by somatic cell nuclear transfer (SCNT) had identical genetic background except for cytoplasmic transmission molecules, which originated from recipient oocytes. To identify the SCNT calves generated from 3 nucleus donors, genotypes for 16 microsatellite (MS) markers and mitochondrial DNA (mtDNA) polymorphisms were analyzed. Using MS genotypes, the parentage test results were not only classified as the single donor-derived calves but mtDNA sequence variations might also discriminated all SCNT calves within the donor-derived families. Comparing MS genotypes, AI-derived progenies were easily discriminated from each other. However, those genotypes could not supply the useful information for identifying the SCNT calves produced from each donor. Informative sequence variations were detected in several regions including D-loop, 12 S rDNA, and ND5 genes. About 19 nucleotide substitutions found within D-loop allowed individual identification for most SCNT-derived progeny except for 5 individuals. However, further investigation on 12 S rDNA and ND5 genes provided the useful polymorphic information for those 5 individuals. Although the experiment had been carried out to produce SCNT calves without previous investigation of mtDNA polymorphism, polymorphic mtDNA sequences provided interesting information that discriminated individuals, even those from the same donor cells. In addition, we could distinguish the 2nd generations produced by AI combinations using SCNT donors and SCNT progeny as the dams and/or the sires when combined molecular data obtained from MS genotypes and mtDNA polymorphisms were derived. These results suggested that mtDNA polymorphisms might supply the critical information for identification and traceability for SCNT-derived calves when combined with MS data. This work received grant support from the Agenda Program (No. 200901FHT010305191), Rural Development Administration, Republic of Korea.