1989 Fiscal Year Final Research Report Summary
A MUTATION DETECTION SYSTEM FOR MAMMALS AT INDIVIDUAL LEVEL
| Project/Area Number |
63580168
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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 |
放射線5生物学
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| Research Institution | Nihon University, College of Agriculture and veterinary Medicine |
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Principal Investigator |
INOUE Tadashi Nihon Univ., Col. of Agric. and Veter. Med. Associate Professor, 農獣医学部, 助教授 (90124213)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Mutation / Mouse / Lambda phage |
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| Research Abstract |
Quantitative assay for mutation induction in mammals at individual level is of importance to evaluate the effect on humans of environmental mutageiis and carcidogens as well as genotoxic agents, which have been recently used in modern living. Over hundreds thousands of mice are required, however, to carry out experiments for quantitation of mutation frequencies. We therefore tried to develop an assay system in which only a small number of animals are required for the detection of mutation at the individual level.
The system has been constructed by combination of phage genetics and transgenic mouse system. As a marker gene, supF gene of E. coli was introduced into lambda phage EMBL3. When mixed with their proteins, the DNA of lambda phage can be packaged in vitro to yield lambda phage particles. Mutation in supF gene in the lambda phages is easily detected by using a proper E. coli as an indicator cells.
Purified DNA of the lambda phage containing supF was injected into fertilized eggs of mice to establish a transgenic mouse. Presence of the transgene in the developed mice was examined by southern analysis of the DNA extracted from tails using supF gene as a probe. To date, we have obtained several strains of transgenic mice. We are now examining the organization of the transgene in the transgenic mice by restriction analysis. Once a strain with appropriate properties has established, DNA from sperms from the mice is subjected to the in vitro packaging system to analyze the mutation in supF gene occurred in vivo.
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