The effects of ADH3p overexpression on the mitochondrial morphology in Saccharomyces cerevisiae

• Main Authors: Chia-Hung Ku, 古佳弘
• Other Authors: Ming-Kai Chen
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/81316963710897552039

碩士 === 淡江大學 === 生命科學研究所碩士班 === 98 === The main purpose of this research is to study the effect of over-expression of ADH3p on mitochondrial morphology in Saccharomyces cerevisiae. We constructed two plasmids; one contains wild-type ADH3 sequence in pYES2; another contains ADH3G211A which has a point mutation in ADH3 sequence. We transformed these plasmids and control plasmid without any extra sequence into Saccharomyces cerevisiae. Then we induced ADH3p over-expression with galactose. After induction, we extracted total protein from yeast cells by a sonifier. To quantify the ADH3p and ADH3pG211A being induced, SDS-PAGE was carried out and specific activity was measured. Then stain yeast mitochondria by MitoTracker Green FM and observe the morphological changes under fluorescence microscopy. We find the proportion of network mitochondria is decreased, and that of particulate mitochondria is increased. We also used FACS to measure fluorescence strength. The fluorescence strength of the ADH3p over-expressing strain is stronger than the control.