PROTEIN EXPRESSIONS IN MITOCHONDRIA ISOLATE FROM KALANCHOË DAIGREEMONTIANA

Abstract

In this study, we isolated and purified mitochondria from leaves of Kalanchoë daigreemontiana by using Percoll gradients. The purified mitochondrial were used to isolate protein and mitochondrial protein were determined. Mitochondrial purified protein was extracted in lysis buffer containing protease inhibitor mix, followed by precipitation with ice-cold acetone. Two-dimensional electrophoresis (2DE) was used to investigate and identify mitochondrial protein expression of K. daigreemontiana. This investigation focused on proteins in the isoelectric point range of 3-10 and molecular weight range from 14.4 to 97.4 kDa. Staining of 2 DE gel with Sypro Ruby revealed more than 150 major protein spots from mitochondria. The 2DE maps of K. daigreemontiana mitochondrial soluble proteins showed less vertical streaking indicating that well proteins were transferred from strips into the gel.