Title Page
Contents
I. Introduction 8
1. Trend of longevity research 8
2. Fission yeast as a model for longevity research 10
3. Longevity-related pathways and genes studied in fission yeast 12
4. Purpose 14
II. Materials and Methods 15
1. Strains and Medium 15
2. Microarray 16
2.1. Sampling 16
2.2. Genomic DNA purification 17
2.3. Barcode amplification by polymerase chain reaction (PCR) 19
2.4. Chip hybridization 21
3. Solid medium inoculation 23
4. Gene Ontology (GO) analysis 25
5. Growth curve measurement 25
6. Cell morphology observation 25
7. Measurement of lifespan change according to glucose concentration 26
8. Stress-sensitivity measurement 26
9. Nuclear fragmentation measurement by stress-inducing substances 27
III. Results 28
1. Long-lived mutants screening using fission yeast haploid deletion mutant library 28
2. Individual viability verification of long-lived candidates 31
3. Gene Ontology analysis of long-lived mutants 34
4. Most of the long-lived mutants had a lower growth rate 36
5. Analysis of morphological phenotype of long-lived mutants 40
6. Changes in lifespan of △idn1 according to glucose concentration 43
7. △idn1 increased sensitivity to oxidative stress by decreasing NADPH production 47
8. △pca1 is resistant to apoptosis-induced substances 50
IV. Discussion 55
V. Reference 61
국문초록 66
ABSTRACT 68
Table 1. PCR mixture composition for barcode amplification 20
Table 2. PCR protocol 20
Table 3. Primer for PCR amplification of barcode 20
Table 4. Composition of hybridization 22
Table 5. Composition of staining and washing 22
Table 6. Composition for washing 22
Table 7. Gene Ontology analysis of long-lived mutants 35
Figure 1. Two yeast lifespan fields of study 11
Figure 2. Dilution process for solid medium inoculation. 24
Figure 3. Genome-wide screening of fission yeast using haploid deletion... 30
Figure 4. Plate assay was performed to confirm long-lived mutants in YES medium. 33
Figure 5. Growth rate of long-lived mutants of Group 1. 37
Figure 6. Growth rate of long-lived mutants of Group 2. 38
Figure 7. Growth rate of long-lived mutants of Group 3. 39
Figure 8. Morphological phenotype of long-lived mutants. 42
Figure 9. △idn1 has less change in lifespan due to glucose concentration... [이미지참조] 46
Figure 10. △idn1 is sensitive to oxidative stress. [이미지참조] 49
Figure 11. △pca1 is resistant to AgNP induced apoptosis. [이미지참조] 52
Figure 12. △pca1 is resistant to NaAsO₂ induced apoptosis. [이미지참조] 54
Figure 13. The Schematic outline of glucose metabolic pathways related... 60