표제지
Acknowledgement
목차
Abbreviation 8
Abstract 9
Ⅰ. Introduction 10
Ⅱ. Materials and methods 12
Ⅱ. 1. Animals 12
Ⅱ. 2. Isolation of spermatogenic cells 12
Ⅱ. 3. cDNA subtractive hybridization 12
Ⅱ. 4. Human ESCs and ECCs culture 13
Ⅱ. 5. Differentiation of human ES cells 13
Ⅱ. 6. Reverse transcription_PCR (RT-PCR) 14
Ⅱ. 7. Northern blot analysis 15
Ⅱ. 8. Antibody production 15
Ⅱ. 9. Immunohistochemical analysis 16
Ⅲ. Results 17
Ⅲ.1. Identification and expression of Dppa5 in mice 17
Ⅲ.2. Expression of the Dppa5 in humans 18
Ⅳ. Discussion 20
References 23
Abstract in Korean 27
Figure 1. Alignment of nucleotide sequences for mouse pH 34 (X57708), mouse Esg-1 (AK010584) and mouse Dppa5 (AF490349) 31
Figure 2. Alignment of nucleotide sequences for mouse Dppa5 (AF490349) and human Dppa5 (XM_291161) 32
Figure 3. Expression analysis in various mouse cells by RT-PCR 33
Figure 4. Expression analysis of mDppa5 mRNA in various mouse cells by
Northern blot 34
Figure 5. The expression patterns of hDppa5 in various cells analyzed by RT-PCR 35
A. hDppa5 expression in hESCs, HeLa, hECCs, mESCs and STO 35
B. Expression of hDppa5 and hOct4 in hESCs and differentiated
cells from hESC 35
Figure 6. Confirmation of hDppa5 and hOct-4 expression by Northern blot 36
A. Expression of hDppa5 in MEF, STO, mESCs, HEF, HeLa, hECCs and hESCs 36
B. Expression of hOct4 in MEF, STO, mESCs, HEF, HeLa, hECCs, and hESCs 36
Figure 7. Alignment of amino acid sequences for mouse Dppa5 and antibody
against mDppa5 37
Figure 8. Expression of mDppa5 in colonies of mESCs by immunostaining 38