Analysis of Enzymes functioning in meiosis-specific DNA recombination and isolation of their genes.

分析在减数分裂特异性 DNA 重组中发挥作用的酶及其基因的分离。

• 批准号: 02454551
• 负责人: HOTTA Yasuo
• 金额: $ 3.52万
• 依托单位: Nagoya 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: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454551/
• 关键词: Synaptonema Complex; Pairing of Homologs; Recombination of Genes; Specific Promoters; Chiasma; Topoisomerase; Resolvase; Meiosis; トポイソメラ-ゼI; Recombinase; DNA修複; 減数分裂cDNAクロ-ン; DNA組換え; ヒト肝炎ウィルスDNA断片; Hot Spot; 外来DNAの発現; ポリdG・dC; プロトプラスト

During prophase of meiosis, the homologous chromosomes pair and the reciprocal recombination occurs at high frequency (100-1000 times of somatic recombination) . Such recombination occurs at least one point on each paired homologs. Without stable homologous-pairing, genetic recombination, normal segregation of chromosomes, and formation of reproductive cells do not occur, and thus an organism becomes sterile. During the last study-period we have successfully prepared monoclonal antibodies (one IgG and three IgM) against synaptonemal complex (S. C.) isolated from meiotic nuclei. Although all S. C. from yeast to human have practically identical structure suggesting the presence of common components, this antibody reacted only to lily S. C. but not to S. C. obtained from all organisms tested. We have found topoisomerase and resolvase in S. C. and their activities increased during meiotic pr ophase.Meiotic cells and somatic cells have different DNA-recombination enzyme (s) . We have cloned the specific region in human hepatitis virus, shown to alter the nature of virus, into the mutated pBR322 plasmids and measured the recombination frequency in vitro. We have found 2-5 times increase in the frequency with recombination enzymes from both cells in comparison with the insertion of DNA fragments obtained from other region of virus.35S promoter stimulated somatic gene expression but not meiotic ones. We have found that mei2 promoter helps to express genes in higher plant when introduced into meiotic cells. We have cloned 17 meiosis-specific genes and sequenced all of them. The homology search indicated that three of them had significant similarities with the known protein genes but the others were completely new. AS soon as their promoters are identified, we will study their function in meiosis

在减数分裂前期，同源染色体配对和相互重组发生频率很高（100-1000倍于体细胞重组）。这种重组发生在每对同源物上的至少一个点上。如果没有稳定的同源配对，遗传重组、染色体的正常分离和生殖细胞的形成就不会发生，因此生物体就变得不育。在上一个研究期间，我们成功地制备了抗联会复合体（S。C.）的分离自减数分裂核。虽然所有的S。C.从酵母到人的抗体具有几乎相同的结构，表明存在共同的组分，该抗体仅与百合S反应。C.而不是S。C.从所有测试的微生物中获得。在S. C.在减数分裂前期，DNA重组酶活性增强，减数分裂细胞和体细胞具有不同的DNA重组酶。我们已将人肝炎病毒的特异性区域克隆到突变的pBR 322质粒中，并在体外测定了重组频率。我们已经发现2-5倍的频率增加与重组酶从两个细胞中插入的DNA片段从其他区域的病毒。35 S启动子刺激体细胞基因的表达，但不是减数分裂的。我们发现mei 2启动子导入植物减数分裂细胞后有助于基因的表达。我们已经克隆了17个减数分裂特异性基因并对所有基因进行了测序。同源性分析表明，其中3个基因与已知蛋白质基因有很大的相似性，其余为新基因。一旦确定了它们的启动子，我们将研究它们在减数分裂中的功能

项目成果

S.Tabata: "Control of meiosis." Control of cell growth and division.ed.Ishihama,A.and Yoshikawa,H.91-117 (1991)

S.Tabata：“减数分裂的控制。”

A.Higashitani: "ATP-independent strand-transfer protein from murine spermatocytes,spermatids,and spermatozoa." Exptl.Cell Res.186. 317-323 (1990)

A.Higashitani：“来自小鼠精母细胞、精子细胞和精子的 ATP 独立链转移蛋白。”

Tabata, S., Sato, S. and Hotta, S.: "control of meiosis. in Control of cell growth and division." Springer-Verlag. 97-117 (1991)

Tabata, S.、Sato, S. 和 Hotta, S.：“控制减数分裂。控制细胞生长和分裂。”

Higashitani, A., Tamamoto, S., Tabata, S., Oono, K. and Hotta, Y: "D-loop forming activity and general recombinatory activity in extracts from shoot, flower and callus of varieties of rice." Jap. J. Genet.65. 53-64 (1990)

Higashitani, A.、Tamamoto, S.、Tabata, S.、Oono, K. 和 Hotta, Y：“水稻品种芽、花和愈伤组织提取物中的 D 环形成活性和一般重组活性。”

A.Higashitani: "ATPーindependent strand transfer protein from murine spermatocytes,spermatids,and spermatozoa." Exptl.Cell Res.186. 317-323 (1990)

A.Higashitani：“来自小鼠精母细胞、精子细胞和精子的 ATP 独立链转移蛋白。”Exptl.Cell Res.186 (1990)。