The objective of this study was to analyzed pattern of proteins expression abnormally in cloned bovine placenta. TIMP-2 protein whose function is related to extracellular matrix degradation and tissue remodeling processes was one of differentially up-regulated proteins in SCNT placenta. And one of down-regulated protein in SCNT placenta was identified as vimentin protein that is presumed to stabilize the architecture of the cytoplasm. The expression patterns of these proteins were validated by Western blotting. To evaluate how regulatory loci of TIMP-2 and vimentin genes was programmed reprogramming in cloned placenta, the status of DNA methylation in the promoter region of TIMP-2 and vimentin genes was analyzed by sodium Bisulfite mapping. The DNA methylation results showed that there was not difference in methylation pattern of TIMP-2 and vimentin loci between cloned and normal placenta. Histone H3 acetylation state of the nucleosome was analyzed in the cloned placental and normal placenta by Western blotting. A small portion of the protein lysates were subjected to Western blotting with the antibodies against anti acetyl-Histone H3. Overall histone H3 acetylation state of SCNT placenta was significantly higher than those of normal placenta cells. It is postulated that cloned placenta at the end of gestation seems to be unusual in function and morphology of placenta via improper expression of TIMP-2 and vimentin by abnormal acetylation states of cloned genome.

ABSTRACT   INTRODUCTION   MATERIALS AND METHODS    Placental Samples    Extraction of Solubilized Proteins from Bovine Placentae    2-D Gel Electrophoresis, MALDI-TOF MS and Protein Identification    Genomic DNA Isolation and Bisulfite Treatment    PCR Amplification and Sequencing of the Bisulfite-Rreated Genomic DNA    Western Blot Analysis    Statistical Analysis   RESULTS    TIMPS-2 Expression in SCNT Plancenate    Vimentin in SCNT Placentae    Overall Histone H3 Acetylation State of SCNT Placenta   DISCUSSION   REFERENCES

• Hong-Rye Kim(Research Center for Transgenic Cloned Pigs, Chungnam National University)
• Rong-Xun Han(Research Center for Transgenic Cloned Pigs, Chungnam National University)
• Hye-Ran Lee(Research Center for Transgenic Cloned Pigs, Chungnam National University)
• Jong-Taek Yoon(Genetic Engineering Institute, Hankyong National University)
• Hee-Tae Cheong(School of Veterinary Medicine, Kangwon National University)
• Chang-Sik Park(Research Center for Transgenic Cloned Pigs, Chungnam National University)
• Dong-Il Jin(Research Center for Transgenic Cloned Pigs, Chungnam National University) Corresponding author