1993 Fiscal Year Final Research Report Summary
Analysis of Pathogenesis of Chimera Mice Introduced with the Immediate Early Genes of Murine Cytomegalovirus
| Project/Area Number |
04670222
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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 |
Experimental pathology
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| Research Institution | Institute for Developmental Research, Aichi Parefectural Colony |
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Principal Investigator |
TSUTSUI Yoshihiro Institute for Developmental Research, Department of Morphology, Head, 形態学部, 部長 (50073135)
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| Co-Investigator(Kenkyū-buntansha) |
KASHIWAI Akiko Instiatute for Developmental Research, Department of Morphology, Research Assist, 形態学部, 研究助手
KADOTA Chika Institute for Developmental Research, Department of Morphology, Research Associa, 形態学部, 研究員 (80214419)
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| Project Period (FY) |
1992 – 1993
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| Keywords | cytomegalovirus / chimera mice / gene expression |
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| Research Abstract |
Altough we have made best effort to attain the title of projicts, we could not accomplish the final goal of the project. However, we have overcome several steps to approach the goal and developed the techniques for it.
1) We have done the clonning of the immediate early (IE) genes of murine cytomegalovirus (MCMV) into the pACYC184 plasmid vector as a Bam Hl fragment.
2) We have compared the efficiencies of the IE gene expression after conection of the gene to the expression vector and transfection into various cultured cells.
3) The IE genes were introduced into the embryonic stem cells (ES-D3 cells) by electroporation of the gene and the cloned cells which expressed the IE gene were selected in the presence of G418 by assying using immunofluorescence.
4) We have tried to make chimera mice by inroducing the cloned DE cells into the blastocysts of BDF1 mice and returned in the uteri of pseudopregnant ICR mice. Although we returned the about 100 of the ES cells-injected blastocysts into the uteri of 10 ICR mice, we could not detect the IE gene in the tail DNA of two offspring by PCR amplification.
5) We have also tried to make the transgenic mice using the DNA construct which connect the promoter of the IE gene and lacZ gene. We have gotton 4 transgenic mice out of 36 offspring after injection the construct into the ferilized eggs. We are now under way to examine the expression of the lacZ gene.
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