The embryonic genome activation (EGA) is genetically activated states that embryos make the materials such as growth factors for using themselves. EGA is various because they have many materials, different site, different stage, also different species. At this time, transcription factors are expressed. Transcription factors bind to specific DNA region, and regulate the gene expression. Thus, we check the expression of transcription factors, we can know that embryo development is very well or not. The development stages of embryos are basically the stages from fertilization to blastocyst. So, we check the embryos oocyte to blastocyst. In our experiments, we focus the early developmental transcription factors such as Cdx2, Oct4, Sox2, Nanog and E-Cadherin. Above antibody factors showed different expression sites, and there were many differentiated parts from other animal species. In addition, we compared the SCNT and parthenogenetic activation (PA) because these are same methods using electrical activation among the embryo production methods. Our results showed not only similar patterns but also different patterns between pig and mouse. Therefore, we have to investigate that different patterns of transcription factors play a role in pigs, and why occur.

Oct4 and Nanog are well-known transcription factors related with self renewal of embryonic stem cell. In low-dose of Nanog, transcription of oct4 is increased; however, oct4 is down-regulated upon high-dose of Nanog. There is a negative feedback loop between oct4 and Nanog. To identify this regulation, we generated 4 nested sets for mouse oct4 promoter. Luciferase activities of oct4 were declined upon high-dose Nanog in all constructs. The declined effects of oct4 upon high-dose Nanog were moderated with DNMT and HDAC inhibitors (5-AZA-cytidine and trichostatin A) in 3 constructs (1867, 1346, 754). But, one construct (2179) was only sensitive to TSA. Taken together, these effects were also represented in semi-quantitative RT-PCR and Western blotting data. These data suggest that negative regulation of oct4 gene upon high-dose Nanog would be accomplished by DNMT and HDAC. Further, it will be studied whether these constraining molecules bind to CR1-4 region of oct4 promoter upon low- and high-dose of Nanog.

배아 줄기세포는 미분화상태에서 자가 재생을 유지할 수 있다. 자가 재생은 OCT4, SOX2와 NANOG와 같은 많은 인자들이 작용한다. 생쥐 배아 줄기세포에서 OCT4와 SOX2가 Nanog 프로모터에 결합하여 Nanog 유전자의 발현을 촉진한다는 사실은 생쥐 promoter에 관한 정밀분석으로 알려져 있다. 본 연구에서는 인간 Nanog promoter를 정밀 분석하기 위해 연속적인 결손 돌연변이를 가진 promoter-reporter constru