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Fig. 3 | Retrovirology

Fig. 3

From: Reconstruction of a replication-competent ancestral murine endogenous retrovirus-L

Fig. 3

Reverse transcription-dependent ancML replication. a Organization of the ancML construct. Green arrows indicate promoter sequences. NEO: neo gene in reverse orientation relative to ancML transcription. Chevrons indicate the orientation for each ORF (>: forward, <: reverse). A white box indicates a 33nt deletion in gag at position 671. b G418 resistant colonies on 15 cm cell culture plates derived from CHO cells transfected with plasmids expressing ancML, L1.3 or an empty vector. c Quantification of G418 resistant CHO cell colonies following transfection and treatment in the presence or absence AZT. CHO cells were transfected with plasmids expressing a neo gene (NEO), L1.3, ancML or an ancML construct with inactivating mutations in the RT active site (ancML-RTmut). AZT treatment was applied for 2 days before G418 selection. Data are mean ± SD from 3 independent experiments. d PCR amplification of the neo gene in genomic DNA (gDNA) extracted from CHO cells following transfection with a plasmid expressing ancML or an empty vector. A scheme of the PCR amplification strategy is shown on top. The use of template DNA from plasmid or from CHO gDNA as well as a water control is indicated. M: molecular weight ladder

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