2003 Fiscal Year Final Research Report Summary
Similarity and Diversity of Electron Transport Pathways to Nitrogense, Studies on Rhodobacter and Mesorhizobium
| Project/Area Number |
14540594
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
植物生理
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
SAEKI Kazuhiko Osaka University, Graduate School of Science, Associate Professor, 大学院・理学研究科, 助教授 (40201511)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
FUKUYAMA Keiichi Osaka University Graduate School of Science, Professor, 大学院・理学研究科, 教授 (80032283)
|
|---|
| Project Period (FY) |
2002 – 2003
|
| Keywords | RHODOBACTER / FERREDOXIN / ELECTRON-TRANSPORT / NITROGEN-FIXATION / NADH / MEMBANE-PROTEIN-COMPLEX |
|---|
| Research Abstract |
For structural and biochemical analysis of proteins that participate electron transport to nitrogenase, the following studies were carried out on in Rhodobacter capsulatus rnf and nif gene products and Mesorhizobium loti fixABCX products. (1) Affinity chromatography procedures were optimized for successful purification of the proteins under strict anaerobic conditions with dithionite-containing buffer system, using (His)6-tagged model electron carrier protein. (2) Effective purification procedures were established for R. capsulatus NifF protein after expression in Escherichia coli. (3) The M loti fixX gene was over-expressed in E. coli under various conditions including co-expression of GroES-GroEL chaperones, however, the products formed inclusion body. Reconstitution conditions were investigated to make holo-FixX with Fe-S cluster from denatured inclusion bodies. (4) The M loti fixA, fixB, fixC and fixX genes were co-expressed in E. coli, however, most of the product were situated on membranes. (5) For effective exploitation of the genome sequence information of Lotus microsymbiont, Mesorhizobiurn loti MAFF303099, to discover gene functions, we have constructed an ordered and mutually overlapping cosmid library using an IncP broad host range vector. The library consisted of 480 clones to cover approximately 99.6% of the genome with average insert size and overlap of 26.9 and 11.1 kbp, respectively.
|
Research Products
(12 results)
-
-
-
-
-
-
-
-
-
-
-
[Publications] T.Uchiumi, T.Ohwada, M.Itakura, H.Mitsui, N.Nukui, Dawadi, T.Kaneko, S.Tabata, T.Yokoyama, K.Tejima, K.Saeki, H.Omori, M.Hayashi, T.Maekawa, R.Sriprang, Y.Murooka, S.Tajima, K.Simomura, M.Nomura, A.Suzuki, Y.Shimoda, K.Sioya, M.Abe, K.Minamisawa: "Expression Islands Clustered on the Symbiosis Island of the Mesorhizobium loti Genome"J.Bacteriol.. 186. 2439-2448 (2004)
Description
「研究成果報告書概要(欧文)」より
-
