| Project/Area Number |
60870019
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Virology
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| Research Institution | Kyoto University |
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Principal Investigator |
SHIDA Hisatoshi Institute for Virus Research, Kyoto Univ., ウイルス研究所, 助手 (00144395)
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| Co-Investigator(Kenkyū-buntansha) |
SATAKE Masanobu Institute for Virus Research, Kyoto Univ., ウイルス研究所, 助手 (50178688)
ITO Yoshiaki Institute for Virus Research, Kyoto Univ., ウイルス研究所, 教授 (80004612)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥9,200,000 (Direct Cost: ¥9,200,000)
Fiscal Year 1986: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1985: ¥7,200,000 (Direct Cost: ¥7,200,000)
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| Keywords | Recombinant vaccinia virus / Hemagglutinin gene / Adult T cell leukemia / A型封入体遺伝子 / ワクシニアウイルス / 組換えワクチン / ATL |
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| Research Abstract |
To develop a multivalent vector based on vaccinia virus, we studied the vaccinia virus hemagglutinin (HA) gene and the A-type inclusion body (ATI) gene of cowpox virus.
1. We inserted the env gene of adult T cell leukemia virus (HTLV-I) by the two mwthods and examined the modes of their expressions. The obtained results follow. 1) Vaccinia virus containing the env gene in the HA gene can propagates in vtro and in vivo. 2) The recombinant vaccinia virus could be easily isolated by staining the plaques by chicken erythrocytes; the plaques of the wild-type virus were stained red whereas those of the recombinant viruses remained coloress. 3) The HA promoter was as strong as p7.5 promoter which had been used extensively. These results suggest that the HA gene site is suitable for inserting the foreigen genes.
2. The effect of vaccination by these recombinant vaccinia viruses was examined. 1) They elicited a high titer of anti-env antibodies in injected rabbits. 2) After challenge of HTLV-I no HTLV-I antigen-positive lymphocytes could be detected so far (27 weeks after challenge). Thus, the vaccination of the recombinant virus expressing the env gene protected the rabbits from HTLV-I infection.
3. To clone the ATI gene, the library of cowpox DNA fragments generated by digestion with Sal I was constructed. Then, the library was screened by the methods involving the transfection and transient expression of the ATI gene so as to obtain the plasmid harboring the ATI gene. Further analyses revealed the following characters of the ATI gene. 1) The region upstream of the ATI coding frame has the nucleotide sequence similar to the other late promoters of the vaccinia virus. 2) The open reading frame codes for the protein of 150,000dalton.3) Vaccinia virus also has ATI-related protein of M.W. 94,000.
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