Expression constructs used for schistosome transfection.

<p>(A) The mCherry reporter gene was cloned into the BamHI site of the pCI-neo vector (Promega) to make plasmid pEJ1175. DNA oligos used for amplification by PCR or RT-PCR are shown as a forward arrow (a) or reverse arrows (b and c) representing forward oligo oEJ1022 (a), and reverse oligos oEJ1023 (b) and oEJ1019 (c). (B) 2000 base pairs of the Sm23 UAS was used to control expression of the mCherry and the Sm23 genes, These genes were cloned into the 7.4 kb pGBKT7 vector to make plasmid pEJ1116. (C) The N-terminal 133 amino acids of SmMef2 are regulated by the CMV promoter and were cloned to make plasmid (pEJ1181). The N-terminus of SmMef2 contains the DNA binding domain, but not its C-terminal transactivation domain. (D) The wild-type SmMef2, regulated by the CMV promoter, was cloned to make plasmid (pLS068). DNA oligos (d) and (e) are used for detection of SmMef2,133 transcript by qRT-PCR, while oligos (f) and (g) are used for specifically measuring wt SmMef2 transcript in qRT-PCR reactions.