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. 2016 Mar 9;11(3):e0149409. doi: 10.1371/journal.pone.0149409

Fig 3. Construction and identification of human and mouse proIAPP C. elegans vectors.

Fig 3

A. Schematic representation of human-proIAPP and mouse-proIAPP tissue-specific vector constructs. cDNA of human and mouse proIAPP with human and mouse signal peptide, respectively, were cloned into the BamH1 restriction site of the pSX95.77YFP C.elegans vector to generate human and mouse pSX95.77YFP-proIAPP transgenes. Resulting constructs were digested with Sal1, blunt-ended and ligated into Gateway Cassette C.1 upstream of human and mouse coding sequences to create vectors pNG1 and pNG2. Destination vectors pNG1 and pNG2 were LR recombined with pLR22, pLR25 and pLR35 entry clones to generate human proIAPP plasmids: pPR3, pPR4 and pPR5 and mouse proIAPP plasmids: pPR8, pPR9 and pPR10. B. h-proIAPP sequence of construct pSX95.77YFP-prohIAPP with human signal peptide. Preproh-IAPP (1–89) peptide amino acid sequence is shown in black italics. Signal peptide sequence is shown in red. C. m-proIAPP sequence present in construct pSX95.77YFP-promIAPP with mouse signal peptide. Preprom-IAPP peptide amino acid sequence is shown in black italics. Signal peptide sequence is shown in red. Restriction sites used in the generation of these plasmids are underlined. No stop codon was included after each proIAPP sequence.