Pasion S G, Hartigan J A, Kumar V, Biswas D K
Laboratory of Pharmacology, Harvard School of Dental Medicine, Boston, MA.
DNA. 1987 Oct;6(5):419-28. doi: 10.1089/dna.1987.6.419.
A 10.3-kb DNA fragment in the 5'-flanking region of the rat prolactin (rPRL) gene was isolated from F1BGH(1)2C1, a strain of rat pituitary tumor cells (GH cells) that produces prolactin in response to 5-bromodeoxyuridine (BrdU). Following transfection and integration into genomic DNA of recipient mouse L cells, this DNA induced amplification of the adjacent thymidine kinase gene from Herpes simplex virus type 1 (HSV1TK). We confirmed the ability of this "Amplicon" sequence to induce amplification of other linked or unlinked genes in DNA-mediated gene transfer studies. When transferred into the mouse L cells with the 10.3-5'rPRL gene sequence of BrdU-responsive cells, both the human growth hormone and the HSV1TK genes are amplified in response to 5-bromodeoxyuridine. This observation is substantiated by BrdU-induced amplification of the cotransferred bacterial Neo gene. Cotransfection studies reveal that the BrdU-induced amplification capability is associated with a 4-kb DNA sequence in the 5'-flanking region of the rPRL gene of BrdU-responsive cells. These results demonstrate that genes of heterologous origin, linked or unlinked, and selected or unselected, can be coamplified when located within the amplification boundary of the Amplicon sequence.
从F1BGH(1)2C1（一种大鼠垂体瘤细胞系（GH细胞），其在5-溴脱氧尿苷（BrdU）作用下产生催乳素）中分离出大鼠催乳素（rPRL）基因5'-侧翼区的一个10.3 kb DNA片段。将该DNA转染并整合到受体小鼠L细胞的基因组DNA后，它诱导了来自单纯疱疹病毒1型（HSV1TK）的相邻胸苷激酶基因的扩增。在DNA介导的基因转移研究中，我们证实了这个“扩增子”序列诱导其他连锁或非连锁基因扩增的能力。当与BrdU反应性细胞的10.3-5'rPRL基因序列一起转入小鼠L细胞时，人生长激素基因和HSV1TK基因在5-溴脱氧尿苷作用下均被扩增。BrdU诱导共转入的细菌Neo基因的扩增证实了这一观察结果。共转染研究表明，BrdU诱导的扩增能力与BrdU反应性细胞rPRL基因5'-侧翼区的一个4 kb DNA序列有关。这些结果表明，异源起源的基因，无论是连锁还是非连锁、选择还是未选择的，当位于扩增子序列的扩增边界内时，都可以共同扩增。
