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. 2010 Apr 5;38(11):e123. doi: 10.1093/nar/gkq192

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — (A) Schematic representation of the linear plasmids pShuttle-G5, pBK2272-HPRT and the expected recombination product. The neomycin resistance marker genes (neo) are indicated as solid arrows. The Cre open reading frame is indicated as a shaded arrow. The PGK and the tk promoter are indicated as arrowheads. The copies of the β-globin insulator element (INS) are indicated as vertically striped boxes. The positions of the lox2272 and loxP sites are marked by vertical arrows. (B) PCR analysis of DNA isolated from HEK 293 and BHK cells (as indicated) transfected/infected with pBK2272-HPRT plus the indicated plasmids and virus vectors. The 622-bp PCR product is generated from the non-recombined pBK2272-HPRT plasmid. The 421-bp PCR product (derived from the primer pair PGK5/CMVseq.1) is indicative of a recombinase mediated cassette exchange between the PGK-neo cassette and the PGK-HPRT cassette. Phage λ DNA digested with HindIII and EcoRI was used as molecular weight marker.