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국문요약 12

제1장 서론 14

제2장 재료 및 방법 19

2.1. 실험어 채집 19

2.2. Total DNA 추출 19

2.3. 미토콘드리아 DNA의 선택 유전자 영역 증폭 19

2.4. 염기서열 분석 20

2.5. 분자계통학적 분석 20

2.6. AFLP에 의한 유전 다양성 분석 21

2.6.1. DNA 절단 및 adapter ligation 21

2.6.2. PCR 증폭조건 및 전기영동 22

2.6.3. AFLP 결과 분석 방법 22

제3장 결과 24

3.1. 미꾸리과 어류 및 종개과 어류의 cytochrome b 영역 분석 24

3.2. 미꾸리과 어류 및 종개과 어류의 12S rRNA 영역 분석 25

3.3. 미꾸리과 어류 및 종개과 어류의 D-loop 영역 분석 26

3.4. 미꾸리과 및 종개과 어류의 cytochrome b+12S rRNA+D-loop 영역 분석 27

3.5. AFLP에 의한 유전 다양성 분석 28

제4장 고찰 84

4.1. 분자계통학적 분석 84

4.2. AFLP에 의한 유전다양성 분석 86

참고문헌 89

Abstract 99

표목차

Table 1. Sample sites of the cobitidae and Balitoridae fish from Korea and NCBI acession No. of the outgroup fish in this study 30

Table 2. Design of degeneracy primers from Cypriniformes 32

Table 3. Adapters and primer sequences used for AFLP analysis in this study 33

Table 4. Primer pairs selected for AFLP analysis in I. choii 34

Table 5. Empirical base frequencies of the mitochondrial cytochrome b gene sequences in Family Cobitidae and Balitoridae, other related species belonging to Cypriniformes 45

Table 6. Ratio of transitional and transversional substitution(above) and Kimura 2-parameter distance(below) of a mitochondrial DNA cytochrome b gene among Family Cobitidae 46

Table 7. Empirical base frequencies of the mitochondrial 12S rRNA gene sequences in Family Cobitidae and Balitoridae, other related species belonging to Cypriniformes 59

Table 8. Ratio of transitional and transversional substitution(above) and Kimura 2-parameter distance(below) of a mitochondrial DNA 12S rRNA gene among Family Cobitidae and 60

Table 9. Empirical base frequencies of the mitochondrial D-loop(control reason) sequences in Family Cobitidae and Balitoridae, other related species belonging to Cypriniformes 72

Table 10. Ratio of transitional and transversional substitution(above) and Kimura 2-parameter distance(below) of a mitochondrial DNA D-loop among Family Cobitidae and 73

Table 11. Empirical base frequencies of the mitochondrial cytochrome b+12S rRNA+D-loop sequences in Family Cobitidae and Balitoridae, other related species belonging to 76

Table 12. Ratio of transitional and transversional substitution(above) and Kimura 2-parameter distance(below) of a mitochondrial DNA cytochrome b+12S rRNA+D-loop among 77

Table 13. Summary on the detection of AFLP markers in the three populations of I. choii 81

Table 14. Pairwise distance and Fst calculated between three population by AFLP analysis 82

그림목차

Fig. 1. The sample sites of I. choii. St. 1, Gab stream ; St. 2, Baekgok stream ; St. 3, Ji stream. 31

Fig. 2. Multiple sequence alignment of mitochondrial cytochrome b gene from 21 related species belonging to Family Cobitidae, Balitoridae, Cyprinidae. Dots denote identity with 44

Fig. 3. Maximum likelihood tree obtained from mitochondrial cytochrome b gene sequences of 19 ingroup and 2 outgroup species. Numbers at the branching point show 47

Fig. 4. Neighbor-joining tree obtained from mitochondrial cytochrome b gene sequences of 19 ingroup and 2 outgroup species produced using Kimura 2-parameter(Kimura, 1980). 48

Fig. 5. Multiple sequence alignment of mitochondrial 12S rRNA gene from 21 related species belonging to Family Cobitidae, Balitoridae, Cyprinidae. Dots denote identity with the 58

Fig. 6. maximum likelihood tree obtained from mitochondrial 12S rRNA gene sequences of 19 ingroup and 2 outgroup species. Numbers at the branching point show probabilities 61

Fig. 7. Neighbor-joining tree obtained from mitochondrial 12S rRNA gene sequences of 19 ingroup and 2 outgroup species produced using Kimura 2-parameter(Kimura, 1980). 62

Fig. 8. Empirical base frequencies of the mitochondrial D-loop(control reason) sequences in Family Cobitidae and Balitoridae, other related species belonging to Cypriniformes. 71

Fig. 9. maximum likelihood tree obtained from mitochondrial D-loop sequences of 19 ingroup and 2 outgroup species. Numbers at the branching point show probabilities(50% or 74

Fig. 10. Neighbor-joining tree obtained from mitochondrial D-loop sequences of 19 ingroup and 2 outgroup species produced using Kimura 2-parameter(Kimura, 1980). 75

Fig. 11. maximum likelihood and Neighbor-joining tree(NJ/ML) obtained from mitochondrial cytochrome b+12S rRNA+D-loop sequences of 19 ingroup and 2 outgroup species. 78

Fig. 12. Neighbor-joining tree obtained from mitochondrial cytochrome b+12S rRNA+D-loop sequences of 19 ingroup and 2 outgroup species produced using Kimura 2-parameter 79

Fig. 13. AFLP fingerprint patterns from three populations of I. choii generated with a primer combination of E/ACT-M/CAT. 1-15 lane, Gab stream population ; 16-30 lane, Baekgok 80

Fig. 14. Dendrogram showing phylogenetic relationships among three populations based on genetic distance data constructed by the UPGMA clustering method. 1-15 lane, Gab 83

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