Molecular dissection of Penelope transposable element regulatory machinery
Supplementary Figure 1: The 5’end of Penelope-CAT transcripts from the pool of total RNA isolated from the ovaries or carcasses of flies from transgenic strains transformed with constructs Р and С were determined by 5' RACE using nested primers within the CAT sequence (91.98Kb)
Supplementary Figure 2: Sequences of cDNAs from the ovaries or carcasses of flies carrying constructs A, or G showing that unspliced RNA could be detected only in the carcasses of flies with construct A and that intronless RNA could be detected in both the carcasses and ovaries of flies with construct G (13.17Kb)
Arkhipova, Irina R.
Finnegan, David J.
Evgenev, Michael B.
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Penelope-like elements (PLEs) represent a new class of retroelements identified in more than 80 species belonging to at least 10 animal phyla. Penelope isolated from Drosophila virilis is the only known transpositionally active representative of this class. Although the size and structure of the Penelope major transcript has been previously described in both D. virilis and D. melanogaster transgenic strains, the architecture of the Penelope regulatory region remains unknown. In order to determine the localization of presumptive Penelope promoter and enhancer-like elements, segments of the putative Penelope regulatory region were linked to a CAT reporter gene and introduced into D. melanogaster by P-element-mediated transformation. The results obtained using ELISA to measure CAT expression levels and RNA studies, including RT–PCR, suggest that the active Penelope transposon contains an internal promoter similar to the TATA-less promoters of LINEs. The results also suggest that some of the Penelope regulatory sequences control the preferential expression in the ovaries of the adult flies by enhancing expression in the ovary and reducing expression in the carcass. The possible significance of the intron within Penelope for the function and evolution of PLEs, and the effect of Penelope insertions on adjacent genes, are discussed.
© 2008 The Authors. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The definitive version was published in Nucleic Acids Research 36 (2008): 2522-2529, doi:10.1093/nar/gkm1166
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