McBride O W, Rajagopalan M, Hatfield D
J Biol Chem. 1987 Aug 15;262(23):11163-6.
An opal suppressor phosphoserine tRNA gene and pseudogene have been isolated from a human DNA library and sequenced (O'Neill, V., Eden, F., Pratt, K., and Hatfield, D. (1985) J. Biol. Chem. 260, 2501-2508). Southern hybridization of human genomic DNA with an opal suppressor tRNA probe suggested that the gene and pseudogene are present in single copy. In this study, we have determined the chromosome location of the human gene and pseudogene by utilizing a 193-base pair fragment encoding the opal suppressor phosphoserine tRNA gene as probe to examine DNAs isolated from human-rodent somatic cell hybrids that have segregated human chromosomes. These studies show that the probe hybridized with two regions in the human genome; one is located on chromosome 19 and the second on chromosome 22. By comparing the restriction sites within these two regions to those previously determined for the human opal suppressor phosphoserine tRNA gene and pseudogene, we tentatively assigned the gene to chromosome 19 and the pseudogene to chromosome 22. These assignments were confirmed by utilizing a 350-base pair fragment which was isolated from the 5'-flanking region of the human gene as probe. This fragment hybridized only to chromosome 19, demonstrating unequivocally that the opal suppressor phosphoserine tRNA gene is located on chromosome 19. The flanking probe hybridized to a single homologous band in hamster and in mouse DNA to which the gene probe also hybridized, demonstrating that the 5'-flanking region of the opal suppressor tRNA gene is conserved in mammals. Restriction analysis of DNAs obtained from the white blood cells of 10 separate individuals demonstrates that the gene is polymorphic. This study provides two additional markers for the human genome and constitutes only the second set of two tRNA genes assigned to human chromosomes.
已从人类DNA文库中分离出一个乳白抑制子磷酸丝氨酸tRNA基因和假基因并进行了测序（奥尼尔，V.，伊登，F.，普拉特，K.，和哈特菲尔德，D.（1985年）《生物化学杂志》260，2501 - 2508）。用人乳白抑制子tRNA探针与人基因组DNA进行Southern杂交表明，该基因和假基因以单拷贝形式存在。在本研究中，我们通过利用编码乳白抑制子磷酸丝氨酸tRNA基因的193个碱基对片段作为探针，检测从已分离出人类染色体的人 - 啮齿动物体细胞杂种中分离的DNA，确定了人类基因和假基因的染色体定位。这些研究表明，该探针与人类基因组中的两个区域杂交；一个位于19号染色体上，另一个位于22号染色体上。通过将这两个区域内的限制性酶切位点与先前确定的人类乳白抑制子磷酸丝氨酸tRNA基因和假基因的位点进行比较，我们初步将该基因定位于19号染色体，将假基因定位于22号染色体。利用从人类基因的5'侧翼区域分离出的350个碱基对片段作为探针，证实了这些定位。该片段仅与19号染色体杂交，明确表明乳白抑制子磷酸丝氨酸tRNA基因位于19号染色体上。侧翼探针与仓鼠和小鼠DNA中的一条同源带杂交，该基因探针也与这条带杂交，表明乳白抑制子tRNA基因的5'侧翼区域在哺乳动物中是保守的。对来自10个不同个体白细胞的DNA进行限制性分析表明，该基因具有多态性。本研究为人类基因组提供了另外两个标记，并且是仅有的第二组被定位于人类染色体的两个tRNA基因。
