Development of highly efficient homologous recombination system using mouse recombination genes and establishment of mouse strains with gene engineered

利用小鼠重组基因开发高效同源重组系统并建立基因工程小鼠品系

• 批准号: 06454717
• 负责人: MORITA Takashi
• 金额: $ 2.94万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454717/
• 关键词: highly efficient recombination system; Rad51 gene; RecA gene; cell cycle; DNA repair; gene targeting; Dmc1 gene; meiosis; Red51遺伝子

The mouse Rad51 gene is a mammalian homologue of the Escherichia coli recA and yeast RAD51 genes, both of which are involved in homologous recombination and DNA repair in mitosis and meiosis. The expression of mouse Rad51 mRNA was examined in cell-cycle synchronized mouse m5S cells. The Rad51 transcript was observed from late G1 phase through to M phase. During the period of late G1-S-G2 phase, the RAD51 proteins were exclusively observed in nuclei. Activation of T cell and B cell proliferation in spleen by mitogens induced the expression of Rad51 mRNA.By immunohistochemical analyzes, the mouse RAD51 protein was detected in proliferating cells ; spermatogonia in testis, immature T cells in thymus, germinal center cells of the secondary lymphatic nodules of spleen and intestine, follicle cells in ovary and epithelial cells in uterus and intestine. It was also expressed in spermatocytes during the early and mid prophase of meiosis and in resting oocytes before maturation. Thus mouse Rad51 expression is closely related to the state of cell proliferation and is presumably involved in DNA repair coupled with DNA replication as well as meiotic DNA recombination in spermatocytes.

小鼠Rad 51基因是大肠杆菌recA和酵母RAD 51基因的哺乳动物同源物，两者都参与有丝分裂和减数分裂中的同源重组和DNA修复。在细胞周期同步的小鼠m5 S细胞中检测小鼠Rad 51 mRNA的表达。从G1期晚期到M期观察到Rad 51转录本。在G1-S-G2期晚期，RAD 51蛋白仅在细胞核中表达。免疫组化结果显示，小鼠脾脏T细胞和B细胞的增殖可诱导Rad 51 mRNA的表达，小鼠睾丸精原细胞、胸腺未成熟T细胞、脾脏和小肠次级淋巴结的生发中心细胞、卵巢滤泡细胞以及子宫和小肠上皮细胞中均有Rad 51蛋白的表达。在减数分裂早期和中期的精母细胞以及成熟前的休眠卵母细胞中也有表达。因此，小鼠Rad 51的表达与细胞增殖状态密切相关，并可能参与DNA修复与DNA复制以及精母细胞减数分裂DNA重组。

项目成果

T. Habu, et al.: "The mouse and human homologues of DMCq, the yeast meiosis-specific homologous recombination gene, have a common unique form of exon-skipped transcript in meiosis." Nucleic Acids Res.24(in press). (1996)

T. Habu 等人：“DMCq（酵母减数分裂特异性同源重组基因）的小鼠和人类同源物在减数分裂中具有共同的、独特的外显子跳跃转录本形式。”

Kato, T., et al.: "Cloning of a marsupial DNA photolyase gene and the lack of related nucleotide sequences in placental mammals." Nucleic Acids Res.22,. 4119-4124 (1995)

Kato, T. 等人：“有袋动物 DNA 光裂合酶基因的克隆以及胎盘哺乳动物中相关核苷酸序列的缺乏。”

Yagi, H., et al.: "Regulation of the Mouse Histone H2A. X gene Promoter by the Transcription Factor E2F and CCAAT Binding Protein" J. Biol. Chem.270. 18759-19765 (1995)

Yagi, H. 等人：“转录因子 E2F 和 CCAAT 结合蛋白对小鼠组蛋白 H2A.X 基因启动子的调节”J. Biol。

Matsuda, Y. et al.: "Chromosome mapping of the mouse homologue of DMC1, the yeast meiosis-specific homologous recombination gene" Chromosome Res.(in press). (1996)

Matsuda, Y. 等人：“DMC1（酵母减数分裂特异性同源重组基因）的小鼠同源物的染色体作图”Chromosome Res.（出版中）。

Takahashi,E.et al.: "Chromosome mappings of the human (RECA) and mouse (Reca) Homologs of the yeast RAD51 and E coli recA genes to human (15n15.1) and mouse (2F1) chromosomes by direct R-bnading fluorescence in situ hybridization." Genomics. 19. 376-378 (

Takahashi,E.et al.：“通过直接 R-bonding 将酵母 RAD51 和大肠杆菌 recA 基因的人类 (RECA) 和小鼠 (Reca) 同源物与人类 (15n15.1) 和小鼠 (2F1) 染色体进行染色体映射