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Fig. 2 | Mobile DNA

Fig. 2

From: Deciphering fact from artifact when using reporter assays to investigate the roles of host factors on L1 retrotransposition

Fig. 2

Schematic of L1 reporter plasmids. All reporters contain a full-length L1 element with 5′ and 3′ UTRs (orange), ORF1 (pink), intergenic region (gray), ORF2 (blue) and a retrotransposition reporter (yellow) interrupted by an artificial intron (purple) with splice donor (SD) and acceptor (SA) sites. In JM101, L1 is driven by the CMV promoter (green), and in pYX017 by the hybrid CAG promoter (green). pYX014 contains only the native L1 promoter in the 5′UTR, and pYX015 is identical to pYX014 except for two missense mutations (R261A/R262A) [38] in ORF1p, rendering pYX015 incompetent for retrotransposition. The reporter in JM101 is an mneo cassette driven by the SV40 promoter (green) located within the 3′ UTR. The pYX017, pYX014 and pYX015 constructs contain a Firefly luciferase reporter (Fluc), also driven by SV40 (green), as well as a gene for Renilla luciferase (Rluc; aqua) driven by the HSV-TK promoter (green)

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ISSN: 1759-8753