Title page
ACKNOWLEDGEMENT
Contents
Abstract 10
I. Introduction 11
II. Materials and methods 14
II. 1. Subject 14
II. 2. Clinical and biochemical analysis 14
II. 3. Restriction fragment length polymorphism (RFLP) analysis 14
II. 4. Direct sequencing analysis 15
II. 5. TaqMan analysis 18
II. 6. Association test 18
II. 7. Haplotype analysi 21
III. Results 22
III. 1. Clinical and biochemical characteristics 22
III. 2. Genotyping analysis and association test for SNPs in INSR gene with RFLP assay 25
III. 3. Screening of candidate SNPs in INSR gene 28
III. 4. Genotyping analysis and association test for SNPs in INSR gene with TaqMan analysis 28
III. 5. Haplotype analysis for INSR gene 31
III. 6. Screening of candidate SNPs in VEGF gene 33
III. 7. Genotyping analysis and association test for SNPs in VEGF gene with TaqMan assay 35
III. 8. Haplotype analysis for VEGF gene 37
IV. Discussion 39
V. Conclusions 42
Reference 43
Summary in Korean 49
Table 1. Sequences of 25 primer sets for candidate SNP screening in INSR gene. 16
Table 2. Sequences of 12 primer sets for candidate SNP screening in VEGF gene. 17
Table 3. Sequences of primers and probes for large scale genotyping of SNPs in INSR gene with TaqMan assay. 19
Table 4. Sequences of primers and probes for large scale genotyping of SNPs in VEGF gene with TaqMan assay. 20
Table 5. A. Clinical and biochemical characteristics of subjects for RFLP analysis. B. Clinical and biochemical characteristics of subjects for TaqManan analysis. 23
Table 6. Genotyping analysis and association test for C/T polymorphism of exon 17 of INSR gene in PCOS patient group (n=174) and control group (n=93) by RFLP analysis. 26
Table 7. Genotyping analysis and association test for candidate SNPs in INSR gene. 30
Table 8. Haplotype analysis for candidate SNPs in INSR gene. 32
Table 9. Genotyping analysis and association test for candidate SNPs in VEGF gene. 36
Table 10. Haplotype analysis for candidate SNPs in VEGF gene. 38
Figure 1. The proportion of subjects which shown symptoms of PCOS and enrolled for RFLP analysis. 24
Figure 2. (A) The structure of amplified region in the exon 17 of INSR gene. An arrow indicates the restriction site of PmlI. The site of the His 1085 C/T polymorphism is marked 27
Figure 3. Map of INSR on chromosome 19p13.3-19p13.2 (177 kb). Black blocks indicate coding exons and white blocks indicate 5' and 3' UTR. 9 sites of SNPs marked with asterisks 29
Figure 4. Map of VEGF on chromosome 6p12 (14 kb). Black blocks indicate coding exons and white blocks indicate 5' and 3' UTR. 10 sites of SNPs marked with asterisks (*) were 34