一个包含XRCC5（Ku80）DNA修复基因的酵母人工染色体连续克隆系，以及通过酵母人工染色体原生质体融合对缺陷细胞进行互补。

A YAC contig encompassing the XRCC5 (Ku80) DNA repair gene and complementation of defective cells by YAC protoplast fusion.

作者信息

Blunt T, Taccioli G E, Priestley A, Hafezparast M, McMillan T, Liu J, Cole C C, White J, Alt F W, Jackson S P

机构信息

MRC Cell Mutation Unit, University of Sussex, Brighton, United Kingdom.

出版信息

Genomics. 1995 Nov 20;30(2):320-8. doi: 10.1006/geno.1995.9871.

DOI:10.1006/geno.1995.9871

PMID:8586433

Abstract

The Chinese hamster ovary xrs mutants are sensitive to ionizing radiation, defective in DNA double-strand break rejoining, and unable to carry out V(D)J recombination effectively. Recently, the gene defective in these mutants, XRCC5, has been shown to encode Ku80, a component of the Ku protein and DNA-dependent protein kinase. We present here a YAC contig involving 25 YACs mapping to the region 2q33-q34, which encompasses the XRCC5 gene. Eight new markers for this region of chromosome 2 are identified. YACs encoding the Ku80 gene were transferred to xrs cells by protoplast fusion, and complementation of all the defective phenotypes has been obtained with two YACs. We discuss the advantages and disadvantages of this approach as a strategy for cloning human genes complementing defective rodent cell lines.

摘要

中国仓鼠卵巢细胞xrs突变体对电离辐射敏感，在DNA双链断裂重新连接方面存在缺陷，并且无法有效地进行V(D)J重组。最近，这些突变体中存在缺陷的基因XRCC5已被证明编码Ku80，Ku蛋白和DNA依赖性蛋白激酶的一个组成部分。我们在此展示了一个由25个酵母人工染色体（YAC）组成的重叠群，这些YAC定位到2q33 - q34区域，该区域包含XRCC5基因。确定了2号染色体这个区域的8个新标记。通过原生质体融合将编码Ku80基因的YAC转移到xrs细胞中，并且用两个YAC获得了所有缺陷表型的互补。我们讨论了这种方法作为克隆互补缺陷啮齿动物细胞系的人类基因策略的优缺点。