The <i>bzpR</i> 5′ UTR sequence and plasmids used to test translational suppression.
2013-02-20T08:07:24Z (GMT) by
<p><b>A</b>. A ‘to scale’ schematic of the seven uORFs within the <i>bzpR</i> 5′ UTR, color coded to correspond with panels B and C. <b>B</b>. The <i>bzpR</i> 5′ UTR sequence and the first nine codons (blue) of the coding region. The 5′ ends of the three primers, bzr-9, bzr-10, and bzr-5, used in RT-PCR to localize the transcriptional start site, are indicated. Transcription likely starts between the two underlined regions. The sequence shown was fused to the GFP gene in pbzpR-17. The italicized region, consisting only of the ribosome binding site and the first 9 codons, is the no-UTR control fused to the GFP and RFP genes in pbzpR-14 and to the RFP gene in pbzpR-17, -53, and -54. The larger font T and A in the first and last uORFs represent the fusion junction in the internal deletion constructs (pbzpR-53 and -54). <b>C</b>. Schematic diagram of the plasmids used to demonstrate suppression of translation by the <i>bzpR</i> 5′ UTR. A15 represents the actin 15 promoter driving transcription of each fusion. Thin blue lines represent the remaining portions of the plasmids, including transcriptional terminators and selectable drug resistance genes.</p>