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Fig. 5 | Parasites & Vectors

Fig. 5

From: Transcription of Leishmania major U2 small nuclear RNA gene is directed by extragenic sequences located within a tRNA-like and a tRNA-Ala gene

Fig. 5

Transcription of the tRNA-like sequence associated to the U2 snRNA gene. a Northern blot analyses with specific probes to detect the tRNA-Ala (left image), tRNA-like (middle image) and U2 snRNA (right image). b Mapping of the 5' ends of tRNA-like molecules in wild type promastigotes of L. major. Total RNA was subjected to primer extension analysis with oligonucleotide 5'LmjU2DcajaB, and the products were separated on an 8 % polyacrylamide denaturing gel (Lane PE) together with a DNA sequencing ladder obtained with the same oligonucleotide (Lanes G, C, T and A). The primer extension products are marked with asterisks; the red asterisk denotes the most abundant product. Partial sequences of the tRNA-like are shown on the left, indicating with arrows the positions of the 5' termini and the expected size of tRNA-like molecules. c Sequence of the tRNA-like, which is boxed with dotted lines (orange box with an arrow indicating the direction of transcription). Primer extension products are marked with arrows, indicating the length of the transcript. The most abundant primer extension product was considered as the TSS (C, 109 nt). Boxes A and B are colored in green and yellow, respectively. The tRNA-like transcription stop signal is shown in red. The location of the 5'LmjU2DcajaB oligonucleotide is indicated. The first eleven nucleotides of the U2 snRNA gene are also shown. Sequence numbers are relative to the TSS (+1) from the U2 snRNA. To facilitate the analysis of the results, the sequence is shown as presented in the TriTrypDB database (which is the reverse-complement of the sequence shown in Fig. 1)

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