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DNA sequence responsible for the amplification of adjacent genes
S G Pasion1, J A Hartigan, V Kumar
1Laboratory of Pharmacology, Harvard School of Dental Medicine, Boston, MA.
DNA (Mary Ann Liebert, Inc.)
|October 1, 1987
Summary
A novel DNA sequence, termed "Amplicon," from rat prolactin gene regulatory regions drives gene amplification in response to 5-bromodeoxyuridine (BrdU). This Amplicon facilitates co-amplification of linked or unlinked genes, offering new tools for genetic engineering.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- Rat pituitary tumor cells (GH cells) produce prolactin when exposed to 5-bromodeoxyuridine (BrdU).
- Gene amplification is a crucial mechanism for increasing gene expression and is utilized in various biotechnological applications.
Purpose of the Study:
- To identify and characterize DNA sequences responsible for BrdU-induced gene amplification.
- To investigate the potential of such sequences to drive co-amplification of heterologous genes.
Main Methods:
- Isolation of a 10.3-kb DNA fragment from the 5 eal-flanking region of the rat prolactin (rPRL) gene.
- Transfection of this fragment into mouse L cells and assessment of its effect on adjacent and co-transfected genes (Herpes simplex virus thymidine kinase [HSV1TK], human growth hormone, bacterial Neo gene).
- DNA-mediated gene transfer studies to confirm amplification capabilities.
Main Results:
- The 10.3-kb DNA fragment, named "Amplicon," induced amplification of the adjacent HSV1TK gene in mouse L cells.
- The Amplicon sequence mediated BrdU-induced co-amplification of linked and unlinked genes, including HSV1TK, human growth hormone, and the Neo gene.
- A 4-kb sub-sequence within the Amplicon was identified as responsible for the BrdU-induced amplification capability.
Conclusions:
- The identified Amplicon sequence acts as a potent enhancer of gene amplification in response to BrdU.
- This discovery provides a novel mechanism for controlling gene amplification and offers potential for applications in genetic engineering and therapeutic development.