| Project/Area Number |
11670267
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Bacteriology (including Mycology)
|
|---|
| Research Institution | University of Shizuoka |
|---|
Principal Investigator |
MASUZAWA Toshiyuki University of Shizuoka, Microbiology, Associate Professor, 薬学部, 助教授 (10181645)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MIYAKE Masaki University of Shizuoka, Microbiology, Assistant Professor, 薬学部, 講師 (00295560)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Project Status |
Completed (Fiscal Year 2000)
|
| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | Lyme disease / Borrelia / Adhesion / Galactosylceramide / Borrelia burgdorferi / glycosphingolipid / GAPDH / Vaccine |
|---|
| Research Abstract |
Glyceraldehyde-3-phosphate dehydrogenase (gapdh) homologue gene, open reading flame about 1020 base pair was cloned into pMAL-c2 plasmid vector and transformed into E.coli JM109. The transformant express ca 80kDa GAPDH protein fused with maltose binding protein (MBP). The recombinant fusion purified with amylose resin was subject to SDS-PAGE and transferred onto PVDF membrane. The recombinant protein was specifically bound with galactosylceramide and was named as glycosphingolipid binding protein (Gbp37). The recombinant Gbp37 fusion protein inhibited the binding of Borrelia cells to CHO-K1 cells at dose 200ug/mL.From immuno electron microscpic observation, anti-Gbp37 sera reacted with antigen expressed borrelia cell surface. However, borrelia cells pretreated with surfactant to disrupt the outer membrane did not reacted with the antisera. The observation indicated Gbp37 located borrelia cell surface. Mice immunized with recombinant protein were challenged with live borrelia cells into footpad. The mice were partially protected from borrelia infection, but were not protect completely. The anti-Gbp37 sera cross-reacted with various borrelia strains belonging to variable borrelia spp. The facts indicated that GAPDH homologue (Gbp37) of Borrelia was one of adhesion molecule expressed cell surface, bound with glycosphingolipid and elicited protective immunity.
|
