2002 Fiscal Year Final Research Report Summary
Studies on superoxide dismutase and oxidative stress response in fishes
Project/Area Number |
12660186
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Fisheries chemistry
|
Research Institution | Nagasaki University |
Principal Investigator |
OSATOMI Kiyoshi Faculty of Fisheries Associate Professor, 水産学部, 助教授 (40253702)
|
Co-Investigator(Kenkyū-buntansha) |
HARA Kenji Faculty of Fisheries Professor, 水産学部, 教授 (10039737)
ISHIHARA Tadashi Faculty of Fisheries Professor, 水産学部, 教授 (40039722)
|
Project Period (FY) |
2000 – 2002
|
Keywords | Superoxide dismutase / SOD / Fish / Purification / Cloning / Fish disease / Bacterial infection / Oxidative stress |
Research Abstract |
Cu, Zn-superoxide dismutase (Cu, Zn-SOD) activities were compared in Japanese flounder Paralichthys olivaceus for four different kinds of bacterial infections. A significant increase in Cu, Zn-SOD levels was recorded in hepato-pancreas at the initial stages of Edwardsiella tarda (E. tarda) infection. It was suggested that the remarkable change of Cu, Zn-SOD activities was a characteristic phenomenon of E. tarda. Cu, Zn-SOD and Mn-SOD were purified to homogeneity from Japanese flounder hepato-pancreas. On SDS-PAGE, the purified Cu, Zn-SOD and Mn-SOD gave single protein bands with molecular mass of 17.8 kDa and 26 kDa under reducing conditions, respectively. The N-terminal ammo acid sequences were determined for 25 amino acid residues (Cu, Zn-SOD) and 19 residues (Mn-SOD), the sequences were compared with other SODs. Reverse transcription-polymerase chain reaction of RNA from Japanese flounder hepato-pancreas was performed using primers designed according to the amino acid sequences determined, and we determined primary structure of Japanese flounder Cu, Zn-SOD and Mn-SOD. Furthermore, recombinant Cu, Zn-SOD (rCu, Zn-SOD) was expressed in Escherichia coli and was recovered as an active form.
|
Research Products
(5 results)