Title Page
ACKNOWLEDGEMENT
Contents
ABBREVIATION 9
ABSTRACT 10
I. INTRODUCTION 11
II. MATERIALS and METHODS 14
II.1. Cell culture and tissue samples 14
II.2. RNA extraction 14
II.3. Suppression subtractive hybridization (SSH) 15
II.4. Analysis of subtraction efficiency 15
II.5. Cloning and sequencing 16
II.6. Bioinformatics analysis 16
II.7. Reverse Transcription-PCR analysis 16
II.8. Quantitative Real-Time PCR analysis 17
II.9. HeLa cell synchronization 17
III. RESULT 18
III.1. Suppression subtractive hybridization (SSH) 18
III.2. Classification of subtracted genes 20
III.3. Gene expression of known genes in various cell lines 27
III.4. Gene expression of known genes in tissue samples 31
III.5. Gene expression of novel genes in cell lines and tissue samples 33
III.6. Gene expression of HERV-HX2 36
III.7. HERV-HX2 expression during the cell cycle 36
IV. DISCUSSION 41
V. CONCLUSION 46
VI. REFERENCES 47
SUMMARY in Korean 54
Table 1. Summary of subtracted cDNA category 21
Table 2. Summary of Database Comparison Results of Known genes Matched to Genbank/NCBI Database Sequences 23
Table 3. Sequences of primer pairs for RT-PCR analysis 29
Table 4. Sequences of primer pairs for HERV-HX2 37
Figure 1. Subtraction efficiency of subtracted cDNAs 19
Figure 2. Functional classification of 116 differentially-expressed known genes 22
Figure 3. Differential expression of known genes by RT-PCR 28
Figure 4. RT-PCR analysis of known genes in various cell lines 30
Figure 5. RT-PCR analysis of known genes in lung tissues 32
Figure 6. RT-PCR analysis of novel genes in various cell lines 34
Figure 7. RT-PCR analysis of known genes in lung tissues 35
Figure 8. Expression pattern of HERV-HX2 in several cell lines 38
Figure 9. Expression pattern of HERV-HX2 in human ES cells and differentiated cells 39
Figure 10. Expression pattern of HERV-HX2 in synchronized HeLa cell 40