Umbricht C B, Griffin C A, Hawkins A L, Grzeschik K H, O'Connell P, Leach R, Green E D, Kelly T J
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Genomics. 1994 Mar 15;20(2):249-57. doi: 10.1006/geno.1994.1161.
Human replication protein A (RPA) is a three-subunit protein that plays a central role in eukaryotic DNA replication, homologous recombination, and excision repair. We have previously reported the cloning and bacterial overexpression of the three RPA genes and have mapped them to chromosome 1 (RPA2), chromosome 7 (RPA3), and chromosome 17 (RPA1). We have now obtained yeast strains with artificial chromosomes carrying the three human RPA genes and report the more detailed genomic mapping of RPA. RPA1 was mapped to chromosome 17p13.3 using a combination of PCR amplification of somatic cell hybrids and radiation hybrids containing chromosome 17 fragments. RPA2 was mapped to chromosome 1p35 by PCR amplification of somatic cell hybrids of chromosome 1 and by fluorescence in situ hybridization. RPA3 was mapped to chromosome 7p22 by Southern analysis and PCR amplification of somatic cell hybrids of chromosome 7 as well as fluorescence in situ hybridization. Since RPA is an essential component of major metabolic events affecting DNA, the physical mapping of the genes for it may help elucidate the biochemical basis of genetic disorders involving DNA metabolism.
人类复制蛋白A(RPA)是一种由三个亚基组成的蛋白质,在真核生物DNA复制、同源重组和切除修复过程中发挥核心作用。我们之前报道了三个RPA基因的克隆及在细菌中的过量表达,并将它们定位到1号染色体(RPA2)、7号染色体(RPA3)和17号染色体(RPA1)上。现在我们获得了携带这三个人类RPA基因的人工染色体酵母菌株,并报道了RPA更详细的基因组定位。利用含有17号染色体片段的体细胞杂种和辐射杂种进行PCR扩增相结合的方法,将RPA1定位到17p13.3。通过对1号染色体的体细胞杂种进行PCR扩增以及荧光原位杂交,将RPA2定位到1p35。通过对7号染色体的体细胞杂种进行Southern分析和PCR扩增以及荧光原位杂交,将RPA3定位到7p22。由于RPA是影响DNA的主要代谢事件的重要组成部分,对其基因进行物理定位可能有助于阐明涉及DNA代谢的遗传疾病的生化基础。