XB-IMG-151663
Xenbase Image ID: 151663
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Fig. 14.
Further analysis of human-Xenopus hybrid Adprhl1 sequences in tadpole hearts. The illustrations at the top indicate the composition of six Adprhl1 hybrids tested for over-expression in transgenic tadpole hearts. Four are predominantly Xenopus adprhl1 cDNAs but with small sequences switched to encode human-derived amino acids within the N-terminal region of the recombinant protein. Importantly, two constructs (M, N, Q, S, T, U) merely contain silent nucleotide changes within the 5’-portion of the Xenopus cDNA, such that the translated product remains identical to the endogenous Adprhl1 protein. A, B, E: Four tadpoles that carry the driver Tg[myl7:Gal4] transgene plus a new integration of a N-terminal switched, Tg[UAS:human1-52-Xenopus53-354adprhl1] responder transgene. Ventral view of stage 44 tadpoles (A) and matching fluorescence image (B) shows anti-Adprhl1 (green) and phalloidin stain in the tail (red). Detail view of the hearts (E) shows abundant hybrid Adprhl1. C, D, F: Tadpoles with the driver plus a Tg[UAS:Xenopus1-52-human53-94-Xenopus95-354 adprhl1] responder do not produce recombinant Adprhl1. G, H, K: The driver plus a complex Tg[UAS:human1-19-Xenopus20-31-human32-52-Xenopus53-354 adprhl1] responder transgene does yield detectable recombinant Adprhl1. I, J, L: The driver plus Tg[UAS:Xenopus1-19-human20-31-Xenopus32-354 adprhl1] does not produce recombinant Adprhl1 (just three positive cells within one heart). M, N, Q: The driver plus the Tg[UAS:Xenopus adprhl1(silent 1-156bp)] responder containing 36 silent nucleotide changes does produce recombinant Adprhl1 (also shown in Fig. 4). O, P, R: Non-transgenic control tadpoles that are siblings to those in (M, N, Q). S, T, U: Additionally, the driver plus the Tg[UAS:Xenopus Adprhl1(silent1-282bp)] responder with 69 silent nucleotide changes also produces recombinant Adprhl1. The Adprhl1-peptide antibody binds at aa residues 249-266. H, heart. Image published in: Smith SJ et al. (2016) © 2016 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |