1993 Fiscal Year Final Research Report Summary
STUDIES ON THE MECHANISM OF SOMATIC MUTATION OF IMMUNOGLOBLIN GENES USING TRANSGENIC MICE.
| Project/Area Number |
04454211
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | NAGOYA CITY UNIVERSITY SCHOOL OF MEDICINE |
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Principal Investigator |
AZUMA Takachika NAGOYA CITY UNIV.SCH.MED., Associate prof., 医学部, 助教授 (00028234)
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| Co-Investigator(Kenkyū-buntansha) |
OKADA Hidechika NAGOYA CITY UNIV.SCH.MED., Prof., 医学部, 教授 (30160683)
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| Project Period (FY) |
1992 – 1993
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| Keywords | Immunoglobulin / Somatic mutation / Transgenic mouse |
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| Research Abstract |
Transgenic mice containing the chloramphenicol acetyl transferase (CAT) gene driven by the immunoglobulin (Ig) V_H promoter and the heavy chain intron enhancer were prepared to increase our ounderstanding of the mechanism responsible for somatic mutation. Hybridomas were prepared after multiple immunization of a transgenic mouse with a thymus dependent antigen. We analyzed mutations in the CAT coding region as well as in the region 5' upstream of the promoter after amplification of DNA using PCR followed by sequencing of cloned DNA.The results of DNA sequencing suggested that mutation occurred only in the CAT gene but not in the promoter or enhancer. The rate of mutetion in the CAT gene was estimated to be 1.6X10^<-5> per base pair per cell division, which was lower limit of the rate for somatic mutation reported for Ig genes. These results suggested that the mutator mechanism can operate in a non-Ig gene provided it is flanked by the V_H promoter and heavy chain intron enhancer.
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