Study for virus replication by using adenovirus expression vector
| Project/Area Number |
06670319
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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 | The University of Tokyo |
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Principal Investigator |
SHIROKI Kazuo The University of Tokyo, Institute of Medical Science, Assistant, 医科学研究所, 助手 (40012744)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1994: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Adenovirus Vector / Poliovirus / Poliovirus receptor / Central nervous system / IRES / ポリオウイルス受容体(PVR) |
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| Research Abstract |
Virus tropisms, such as tissue specificity and host range, are considered to be determined by specific interactions between viral factors and host factors. As for poliovirus, the neurotropism appears to be due to efficiency of IRES function and the host range poliovirus receptor (PVR). Human PVR expression has been detected only in neurons of the CNS of the transgenic mice carrying the human gene for PVR.Thus, PVR expression pattern may determine cell tropism of the virus in the CNS.
To know relationship of IRES function and viral neurotropism, recombinant DNA corresponding to a dicistronic mRNA in which CAT mRNA is for the first cistron and IRES-dependent beta-gal mRNA the second cistron was constructed, and inserted into the adenovirus vector. CAT activity and beta-gal activity in HeLa cells infected with the recombinants were measured at various times after the infection with various moi. Ratios of beta-gal activity to CAT activity were constant in wide ranges of infection time and moi. This indicates that the adnovirus expression system can be used to measure activities of various IRES in infected cells. IRES activities in neurons are now being measured in the infected animals.
To identify the neuron- specific promoter of the human PVR gene, a series of deletions with various length were introduced into the upstream region of human PVR structural gene, and the DNAs were joined to a reporter CAT gene. The reporter genes with truncated upstream sequences were inserted into adenovirus vector that lacked the E1A promoter. CAT activities in HeLa cells infected with recombinant adenoviruses were clearly detected when the reporter gene had about 100 nucleotides of upstream region. The result suggests that the promoter resides in the upstream region of 100 nucleotides long. The promoter activity in neurons are now being investigated in the infected animals.
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Report
(3 results)
Research Products
(6 results)
