DNA sequence responsible for the amplification of adjacent genes.
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.
Pasion
S G
SG
Laboratory of Pharmacology, Harvard School of Dental Medicine, Boston, MA.
Hartigan
J A
JA
Kumar
V
V
Biswas
D K
DK
D000818:Animals; D001973:Bromodeoxyuridine / Q000494:pharmacology; D004274:DNA, Recombinant; D005784:Gene Amplification; D005786:Gene Expression Regulation; D007739:L Cells; D051379:Mice; D010902:Pituitary Gland / Q000502:physiology; D011388:Prolactin / Q000235:genetics*; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
D004274:DNA, Recombinant; D011388:Prolactin; D001973:Bromodeoxyuridine