표제지
목차
국문요약 5
Ⅰ. 서론 7
Ⅱ. 재료 및 방법 9
1. Total RNA 분리 및 cDNA합성과 primer제작 9
2. Primer의 조합과 MTi7의 PCR 10
3. PCR 산물의 cloning 10
4. DNA sequencing과 Data 분석 11
5. 생쥐의 genomic DNA 준비 11
6. 생쥐의 MTi7 inverse PCR primer 제작 12
7. Inverse PCR과 Molecular cloning 12
8. MTi7의 Flanking 유전자의 확인 13
Ⅲ. 결과 14
1. MTi7 element의 분리 14
2. 생쥐 MTi7의 inverse PCR 15
Ⅳ. 고찰 17
Ⅴ. 결론 21
참고문헌 51
영문요약 53
Table 1. BLAST results of MTi7-like elements of each mammalian species 22
Table 2. Homology of MTi7 sequences found in 4 different mammalian species 23
Table 3. BLAST search results for flanking genes of the mouse MTi7 24
Figure 1. Locations of 24bp and 28bp fragments 25
Figure 2. Schematic design for Inverse PCR to find flanking genes outside the mouse MT 27
Figure 3. Mouse MTi7 3' sequence and the design of inverse PCR primers 29
Figure 4. Touch- Down PCR results of MTi7 in four different species by using
fragments and primers in combination. (A) 31
Figure 5. Alignment results of MTi7 33
Figure 6. Homology search of 5′s equences (A), and 3′sequences (B) of MT i7 35
Figure 7. Gradient PCR for MTi7 in each species 37
Figure 8. Bovine MT i7 aligned with the mouse MT i7 39
Figure 9. Porcine MTi7 aligned with the mouse MT i7 41
Figure 10. Rat MTi7 aligned with the mouse MTi7 43
Figure 11. Human MTi7 aligned with the mouse MTi7 45
Figure 12. Electrophores is of 1st and 2nd Inverse PCR products 47
Figure 13. Genomic DNA gradient PCR with F lanking primers 49