Studies for the Development of Recombinant BCG and for the Structure and Function of Secreting Proteins in Mycobacteria
| Project/Area Number |
04670248
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | Aomori University |
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Principal Investigator |
KOBAYASHI Kohmei Aomori Universiy, Faculty of Engineering, Associate Professor, 工学部, 助教授 (70112208)
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| Co-Investigator(Kenkyū-buntansha) |
OHARA Naoya Nagasaki University, School of Dentistry, Assistant, 歯学部, 助手 (70223930)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | M.bovis BCG / AIDS / Malaria / M.avium / alpha-Antigen / MPB70 / Mycobacterium / MPT63 / BCG Tokyo株 / BCG Pasteur株 / α-抗原 |
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| Research Abstract |
The purpose of this study was to develope live recombinant BCG vaccine vehicles into which the genes of antigenic proteins of HIV (for AIDS) or Plasmodium (for malaria) were introduced. DNA containing a B-cell epitope region of HIV-1 p17 gag was fused to a shuttle vector at C-turminal of M.Kansasii alpha-antigen gene. Using this vector, we succeeded to produce the foreign antigen in M.bovis BCG.
The gene for alpha-antigen was isolated from M.avium detected in patients with AIDS.Serological analysis of recombinant M.avium alpha-antigen (A-alpha) indicated the possibility that A-alpha carries at least six B-cell epitopes (Imfection and Immunity, 1993).
To establish more effective expression system, we are interested in studying MPB70 gene in M.bovis BCG substrain Tokyo which excretes large amounts of MPB70. M.bovis BCG substrain Pasteur which do not excrete MPB70 was used as a reference. It was shown by hybridization technique that the N- or C-turminals of the MPB70 gene was retained even in BCG Pasteur (Biomedical Letters, 1993). Next we prepared 1.6kb DNA fragment containing promotor and the gene to examine the process of expression and secretion of MPB70. MPB70 was then produced in the transformant. A variety of shorter DNA fragments containing the MPB70 gene and its upstream have been designed.
To construct the live recombinant BCG vaccine vehicles for malaria, M.bovis BCG was transformed with the hybrid DNA fusing synthetic oligonucleotide based on amino acid sequence of cytotoxic T-lympho cyte (CTL) epitope in the antigenic protein of Plasmodium to MPB64 gene. It was recognized that the foreign antigen was produced in the transformant. However, CTL activities were not induced in mice with the BCG recombinant.
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Report
(4 results)
Research Products
(6 results)
