Hafezparast M, Kaur G P, Zdzienicka M, Athwal R S, Lehmann A R, Jeggo P A
MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton, U.K.
Somat Cell Mol Genet. 1993 Sep;19(5):413-21. doi: 10.1007/BF01233246.
We have previously shown that human chromosome 2 can complement both the radiation sensitivity and the defect in double strand break rejoining characteristic of ionizing radiation (IR) group 5 mutants. A number of human-hamster hybrids containing segments of human chromosome 2 were obtained by microcell transfer into two group 5 mutants. In most, but not all, of these hybrids, the repair defect was complemented by the human chromosomal DNA. Two complementing microcell hybrids were irradiated and fused to XR-V15B, an IR group 5 mutant, to generate further hybrids bearing smaller regions of chromosome 2. All hybrids were examined for complementation of the repair defect. The region of chromosome 2 present was determined using PCR with primers specific for various human genes located on chromosome 2. A complementing hybrid bearing only a small region of chromosome 2 was finally generated. From this analysis we deduced that the XRCC5 gene was tightly linked to the marker, TNP1, which is located in the region 2q35.
我们之前已经表明，人类2号染色体能够弥补电离辐射（IR）5组突变体的辐射敏感性以及双链断裂重新连接缺陷。通过微细胞转移到两个5组突变体中，获得了许多含有人类2号染色体片段的人-仓鼠杂种细胞。在大多数（但并非全部）这些杂种细胞中，修复缺陷被人类染色体DNA所弥补。对两个具有互补作用的微细胞杂种细胞进行辐射，并与IR 5组突变体XR-V15B融合，以产生携带2号染色体更小区域的更多杂种细胞。检查所有杂种细胞的修复缺陷互补情况。使用针对位于2号染色体上的各种人类基因的特异性引物，通过聚合酶链反应（PCR）确定存在的2号染色体区域。最终产生了一个仅携带2号染色体小区域的具有互补作用的杂种细胞。通过该分析，我们推断XRCC5基因与位于2q35区域的标记TNP1紧密连锁。
