Application of replicative lifespan to measure proliferative capacity for the aging study using budding yeast
- 주제(키워드) Chronologicla lifespan , Replicative lifespan
- 발행기관 고려대학교 대학원
- 지도교수 이철구
- 발행년도 2021
- 학위수여년월 2021. 8
- 학위구분 석사
- 학과 대학원 생명공학과
- 세부전공 분자생체공학
- 원문페이지 52 p
- UCI I804:11009-000000251933
- DOI 10.23186/korea.000000251933.11009.0001241
- 본문언어 영어
초록/요약
Saccharomyces cerevisiae, budding yeast, have two different lifespan, chronological lifespan (CLS) and replicative lifespan (RLS). CLS is defined as the length of time cell can survive without nutrient in non-division state, stationary phase. RLS is defined as the number of divisions achieved from parent cell until they stop cell division. Budding yeast can be extended their lifespan by various factors, drug, chemicals, environment, gene and subpopulation. Especially, we measured the lifespan with gene modified budding yeast, tor1△, sfp1△, elo3△, p425, ElO3 OE (Over Expression). TOR1 are highly conserved till higher eukaryotes and related with TORC1 and TORC2 complex, which regulate several cellular processes. These Tor signaling pathway modulates cell growth, cell size, and lifespan in response to nutrient availability, so deletion of Tor signaling pathway reduces cell growth and cell size while increasing lifespan. SFP1 plays important roles in regulating cell size and growth while division and encodes unusual zinc finger transcription factor. Therefore, deletion of SFP1 strain displays slow growth rate and unproper cell size. ELO3 plays a role in synthesis of very long-chain fatty acid (VLCFA) by fatty acid - 3 - elongase 3 and regulates telomere length and life span. Therefore, loss of ELO3 showed reduced telomere length, accelerating chronologic aging, and decreasing RLS. In this study, we used gene-modified budding yeast strains for measuring RLS with the traditional method. We hope that this study will contribute to showing reliavility of RLS measurement and suggesting possibility of correlation with CLS and RLS.
more목차
Abstract…………………………………………………………………………2
Introduction………………………………………………………………………4
1. Drug and chemicals
2. Environment
3. Gene
4. Subpopulation
Materials and Methods………………...……………………….…………….....15
1. Yeast strain and culture condition
2. Measurement of replicative lifespan (RLS)
3. Measurement of Chronological lifespan (CLS) by Colony Forming
Unit (CFU) assay
4. Measurement of CLS by propidium iodide (PI) assay
Results…………………………………………………………………………...20
1. RLS measurement of gene modified yeast strains discovered that
TOR1, SFP1 and ELO3 regulate the lifespan.
2. CLS measurement of gene modified yeast strains showed TOR1, SFP1
and ELO3 regulate the lifespan.
3. RLS and CLS measurement of gene modified yeast strains showed
that two distinctly different methods have meaningful correlation.
4. RLS measurement of gene modified yeast strains showed that
reproductive ability is time-dependent value.
Discussion………….………………………………………….…………………27
References…………………………………………………….…………………34
