Functional Analysis of proteins involved in chromosomal translocations in hematological neoplasms

血液肿瘤染色体易位相关蛋白的功能分析

• 批准号: 12670973
• 负责人: AOKI Katsunori
• 金额: $ 2.37万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670973/
• 关键词: Hematological neoplasm; chromosomal translocation; KO mouse; Translin; 染色体転座; DNA結合蛋白

Translin has been identified as a DNA binding protein that specifically recognizes the consensus sequences motifs at breakpoint junctions in many chromosomal translocations in human lymphoid neoplasms. In the past analysis, many functional features of Translin protein have been characterized. However the physiological function and the association to chromasomal translocations have not been clarified until now. To resolve the problem, we planned the project to make the KO mouse of translin and the associated protein, RP58. Using the known genomic structures and sequences of the both genes, we constructed the target vectors to remove the functional portions of the genes. After the electoroporations of the target vectors to ES cells, we selected the candidate clones using the suitable medium for selection. The genomic DNA of the survived clones were examined by the southern blotting analysis and the objective ES clones were identified. Using these clones, we have finally succeed to make the KO mouse of Translin and RP58 by the conventional method. Now we are energetically making analysis to clarified the physiological functions of both proteins, and would like to prove their involvement to chromasomal translocations.

转译蛋白是一种DNA结合蛋白，它特异性识别人类淋巴肿瘤中许多染色体易位的断点连接处的共有序列基序。在过去的研究中，人们对翻译蛋白的许多功能特性进行了研究。然而，其生理功能及其与染色体易位的关系至今尚未阐明。为了解决这个问题，我们计划制造translin及其相关蛋白RP58的KO小鼠。利用已知的基因组结构和序列，我们构建了靶载体，以去除基因的功能部分。将目的载体转染ES细胞后，用合适的筛选培养基筛选候选克隆。对存活克隆的基因组DNA进行Southern印迹分析，鉴定目的ES克隆。利用这些克隆，我们最终成功地通过常规方法制备了转译蛋白和RP 58的KO小鼠。目前，我们正在积极研究这两种蛋白的生理功能，并希望证明它们参与染色体易位。