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. Author manuscript; available in PMC: 2008 Nov 1.
Published in final edited form as: Mol Biochem Parasitol. 2007 Jul 22;156(1):62–73. doi: 10.1016/j.molbiopara.2007.07.012

Figure 4. Trans-splicing in transgenic mRNAs isolated from embryos transfected with BmATS E1-I1-E2-luc.

Figure 4

Panel A: Schematic diagram of the construct. The elements representing each feature are proportional to their size in the construct (with the exception of the luciferase ORF, which is truncated for clarity). Black boxes indicate exon sequences and lines non-coding sequences (e.g. promoter domains, introns and spacers). The open box represents the luciferase reporter ORF. “SL” indicates the position of the SL1 addition site. “AUG” indicates the first methionine codon in the long open reading frame present in a correctly cis-spliced message, presumably representing the start site of translation. The sequence “AGATGAA” indicates the position of the BmHSP70 TSM homologue found in the first intron of the BmATS gene. Arrowheads over the luciferase gene indicate the position and orientation of the primers used in the trans-splicing assays described in the text. Panel B: SL mediated hemi-nested RT-PCR analyses of RNA prepared from embryos transfected with BmATS E1-I1-E2-luc. Lane A = positive control (RNA extracted from embryos transfected with BmHSP70 E1-I1-E2-luc). Lane B = SL mediated hemi-nested RT-PCRs carried out on mRNA extracted from embryos transfected from BmATS E1-I1-E2-luc carried out in the presence of reverse transcriptase. Lane C = SL mediated hemi-nested RT-PCRs carried out on mRNA extracted from embryos transfected from BmATS E1-I1-E2-luc carried out in the absence of reverse transcriptase.