Basic studies on the introduction of huge DNA molecules into fertilized mouse eggs by homologous recombination between injected fragments.
| Project/Area Number |
05680741
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Laboratory animal science
|
|---|
| Research Institution | Keio University |
|---|
Principal Investigator |
SHIMODA Kouji Keio Univ.School of Medicine, Research Associate, 医学部, 助手 (00129470)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MAEJIMA Kazuyoshi Keio Univ.School of Med.Professor, 医学部, 教授 (70051464)
|
|---|
| Project Period (FY) |
1993 – 1994
|
| Project Status |
Completed (Fiscal Year 1994)
|
| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥1,200,000 (Direct Cost: ¥1,200,000)
|
|---|
| Keywords | Transgenic mice / Homologous recombination / Introduction protocol |
|---|
| Research Abstract |
Production of transgenic mice is now an important tool for the investigation of biomedical research and laboratory animal science. DNA fragments for microinjection are usually used as the minigene construct composed of promoter, coding and terminal sequences, or the genomic fragment encompassed by 5' regulatory and 3' terminal regions. Longer DNA fragment including more informations must have advantages for the precise gene expression in spatial and temporal manner to mimic the original gene expression patterns. Therefore, the method to introduce huge DNA fragments into mouse genome is required for investigation of genes and gene families which are extended for several hundred kilobase long. Our previous research indicated that the fragments injected into fertilized mouse eggs combined to constitute larger fragments by homologous recombination in the overlapping regions of their terminals. In this project we determined the nucleotide sequences in the conjunct regions of homologous and heterogenous recombination by PCR-aid cloning and sequencing. Non-overlapping fragments injected were combined in head to tail orientation, and the deletion and replacement of several nucleotides were observed in the junctions. Overlapping fragments were combined by homologous recombination in their overlapping regions and the replacement of few nucleotide was observed around the junction, however, the deletion of nucleotides was not detected. These results indicated that the reconstitution of large fragments by homologous recombination was a useful protocol for introduction of huge DNA molecules on the mouse genome.
|
Report
(3 results)
Research Products
(9 results)
-
-
-
-
[Publications] Matuo, K., Ikeshima, H., Shimoda, K., Umezawa, J., Hata, J., Maejima, K., Nojima, H.and Takano, T.: "Expression of the rat calmodulin gene II in the central nervous system : a 294-base promoter and 68-base leader segment mediates neuron-specific gene expression in transgenic mice." Mol.Brain Res.20. 9-20 (1993)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Publications] Ikeshima, H., Shimoda, K., Matuo, K., Hata, J., Maejima, K.and Takano, K.: "Spermatocyte-specific transcription by calmodulin gene II promoter in transgenic mice." Mol.Cell.Endocrinol.99. 49-53 (1994)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Publications] Shimoda, K., Ikeshima, H., Matuo, K., Hata, J., Maejima, K.and Takano, T.: "Spatial and temporal regulation of the rat calmodulin gene III directed by a 877-base promoter and 103-base leader segment in the mature and embryonal central nervous system of transgenic mice." Mol.Brain Res.(in press.). (1995)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
-
-
