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Fifty-three human adenovirus serotypes were identified, which are divided into seven subgroups A, B1, B2, C, D, E and F. All types of recombinant adenoviruses have serotype-specific left and right inverted terminal repeats (ITRs). There is evidence that sequences in the ITRs of subgroup C exhibit promoter activity, which in turn might influence the expression of coding sequences that are in close proximity. We investigated whether ITRs from the complete spectrum of adenovirus subgroups show transcriptional activity. We found that ITRs from subgroups A, C and F cloned in a forward orientation display robust transcriptional activity in a cell-type independent manner. In the reverse orientation only subgroup B2 showed transcriptional activity. Unexpectedly, we also found that most ITRs when located upstream of a ubiquitously active promoter drastically reduced reporter gene expression, suggesting that ITRs may have a repressive activity on transcription.
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Journal of General Virology vol. 92 , part 3, pp. 669–674
Supplementary Table S1. Primer sequences used for the amplification of the ITRs of the different adenoviral serotypes, amplification of the PGK promoter and the gene-specific primer for RACE analysis.
Supplementary Methods 1. Cloning of inverted terminal repeats (ITRs) in forward and reverse orientation. 2. Dual luciferase assays to measure transcriptional activity of the ITRs. 3. 5′ RACE
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