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Figure 1 | BMC Molecular Biology

Figure 1

From: The coding/non-coding overlapping architecture of the gene encoding the Drosophila pseudouridine synthase

Figure 1

Molecular structure of mfl mRNAs. (A) Restriction map of the genomic region encompassing the minifly gene (S, Sal I; E, Eco RI; B, Bam HI; N: Not I) [GenBank: AF 097634]. Below, organization of the previously-described 1.8 [GenBank: AF017230] and 2.0 kb [GenBank: AF089837] mfl mRNAs [7], compared with that of the newly identified 2.2 [GenBank: DQ857345] and 1.0 kb mRNA species [GenBank: DQ857346]; note that subsequent releases of the Drosophila genome sequence have revealed that a small intron splits the formerly-designated exon 8 [7] into two moieties, currently indicated as exon 8 and 9. Exonic regions spanned by mfl ORFs are depicted in black. The positions of the intron-encoded H/ACA snoRNA H1 gene [7] and of the DmSnR60 and snm60 isoforms are also shown. (B) The products of the 3'RACE reactions were separated on 2% agarose gels and visualised by ethidium bromide staining. Lane 1, GeneRuler 100 bp DNA ladder (MBI Fermentas). Lane 2, products obtained after amplification with a forward primer derived from exon 5, in combination with the oligo-adaptor reverse primer. Specific fragments of about 800, 650 and 400 bp were obtained; each fragment represents the specific 3' end of a different mfl mRNA, the length of which (in kb) is indicated on the left. Lanes 2 and 3 show negative controls in which no reverse transcriptase or no input RNA were added to the reaction. (C) Northern blot analysis of poly(A)+ RNA extracted from male and female adult flies with a genomic probe derived from exon 12 (probe1: see the genomic map for position). This probe specifically detects the novel 2.2 kb mRNA, most abundant in females, and a large transcript of about 4.4 kb of which the structure remains to be defined. The amount of RNA loaded on each lane was checked by hybridization with a probe derived from αTub84B. The RNA marker I (Roche) was utilised (on the right).

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