| Project/Area Number |
62570206
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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 |
Virology
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| Research Institution | Tokai University School of Medicine |
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Principal Investigator |
IHARA Seiji Tokai University School of Medicine, 医学部, 助教授 (50096202)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Yasushi Tokai University School of Medicine, 医学部, 教授 (60055746)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Human Cytomegalovirus / marker-rescue / DNA replication / 温度感受性変異株 / DNAポリメラーゼ / マーカーレスキュー法 / DNA塩基配列 |
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| Research Abstract |
We have determined the mutation site of a temperature-sensitive mutant of human cytomegalovirus (strain Towne) by marker-rescue assay and DNA sequencing. Analyzed was ts256 which shows a virus specific DNA polymerase minus phenotype. To determine approximately the map position of mutation site of ts256 by marker rescue assay, we first used seven cosmid clones, each carrying an about 40 kb DNA fragment derived from wild type, which cover all the region of HCMV DNA. The ts mutation of ts256 was rescued by pIT212 which covers from 0.22 to 0.37 map unit. Then, by using subcloned fragments derived from pIT212, the position of mutation in pIT212 focused to a narrower region of 5.1 kb from about 0.33 to 0.35 map unit. DNA sequencing of this fragment showed that there existed two open reading frames, one is identical to the putative DNA polymerase gene and the other was unknown gene. A single base change was found within this unknown gene, but not within DNA polymerase gene. This finding suggests that the expression of DNA polymerase activity requires the function of this unknown gene.
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