Functional Analysis of proteins involved in chromosomal translocations in hematological neoplasms

Translin已被鉴定为一种DNA结合蛋白，可特异性识别人类淋巴样肿瘤中许多染色体易位中断点连接处的一致序列基序。在过去的分析中，已经对Translin蛋白的许多功能特征进行了表征。然而，其生理功能及其与染色体易位的关系尚不清楚。为了解决这个问题，我们计划制作translin及其相关蛋白RP58的KO小鼠。利用已知的两种基因的基因组结构和序列，我们构建了目标载体来去除基因的功能部分。将目标载体电泳到胚胎干细胞后，选择合适的培养基进行筛选。用southern blotting法检测存活克隆的基因组DNA，鉴定目标ES克隆。利用这些克隆，我们最终成功地用常规方法制备了Translin和RP58的KO小鼠。目前，我们正在积极进行分析，以澄清这两种蛋白质的生理功能，并希望证明它们与染色体易位有关。

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.