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ARS Home » Research » Publications at this Location » Publication #67076
Title: REPAIR OF DAMAGED TRANSGENES AS A MARKER OF TRANSGENE INTEGRATION.

Author
item Hyman, Paul
item Bondioli, Kenneth
item Wall, Robert

Submitted to: BARC Poster Day
Publication Type: Abstract Only
Publication Acceptance Date: 1/5/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: The ability to identify transgenic embryos before transfer to host mothers would be valuable for increasing the efficiency of producing transgenic animals. One strategy to identify transgenic embryos is to measure activity of the transgene in the embryo. A difficulty with this approach is that transgenes can express before as well as after becoming integrated into the genome. To overcome this problem we have been testing the feasibility of using a damaged transgene containing a single interstrand crosslink caused by psoralen plus UVA treatment. This lesion should prevent expression of the transgene and cannot be repaired until after the transgene is integrated into the genome and undergoes DNA replication. In order to target the crosslink to a specific site in the transgene the psoralen is tethered to an oligonucleotide whose sequence allows it to bind to a specific site in the transgene forming a DNA triple helix. The efficacy of this system is tested using an in vitro cell culture system transfected with B-galactosidase and neomycin resistance expression constructs.