This study was conducted to examine the mRNA expression of apoptosis-related and imprinted genes and methylation pattern of the differentially methylated region (DMR) of H19 gene in day 35 of SCNT pig fetuses. The day 35 of natural mating (control) or cloned (clone) pig fetuses were recovered from uterus. Endometrium from dam and liver from fetus were obtained, respectively. mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. The Bcl-2 mRNA expression in clone was significantly lower than that of control (p<0.05). The mRNA expression of H19 gene in both endometrium and liver was significantly higher in clone than that of control, respectively (p<0.05). The level of IGF-2 mRNA in liver of clone was significantly lower than that of control (p<0.05), whereas the mRNA expression of IGF2-R gene in liver of clone was significantly higher than that of control (p<0.05). The DMR of H19 was lower methylation pattern in clone than that of control. These results suggest that the aberrant mRNA expression of apoptosis-related and imprinted genes and the lower DMR methylation pattern of imprinted gene may be closely related to the inadequate fetal development of cloned fetus.

ABSTRACT   IVTRODUCTION   MATERIALS AND METHODS    Recovery of Control or Cloned Fetuses    Total RNA Extraction and cDNA Synthesis    Quantitative Real-Time PCR    Preparation of Genomic DNA    Sodium bisulfite Genomic Sequencing    Statistical Analysis   RESULTS    Recovery Data of Fetuses from Uterus    mRNA Expression of Apoptosis-Related and Imprinted Genes    DMR Methlation Pattern in H19 Gene   DISCUSSION   REFFTENCES

• Hyun-Ju Jung(Animal Biotechnology Division, National Institute of Animal Science, College of Animal Life Science, Kangwon National University) These autors contributed equally
• Yeoung-Gyu Ko(Animal Biotechnology Division, National Institute of Animal Science) These autors contributed equally
• Seongsoo Hwang(Animal Biotechnology Division, National Institute of Animal Science)
• Gi-Sun Im(Animal Biotechnology Division, National Institute of Animal Science)
• Mi-Rung Park(Animal Biotechnology Division, National Institute of Animal Science)
• Jae-Seok Woo(Animal Biotechnology Division, National Institute of Animal Science)
• Choon-Keum Park(College of Animal Life Science, Kangwon National University)
• Hwan-Hoo Seong(Animal Biotechnology Division, National Institute of Animal Science,) Corresponding author