Figure 1. Four donor plasmids were used to generate transgenic Anopheles stephensi.
nGuy1 and Guy1m were also used in the transient assays described in Table 1. All constructs shown in the figure were flanked by the piggyBac arms to facilitate piggyBac-mediated integration into the An. stephensi genome (Horn et al., 2000). The DsRed fluorescent marker gene under the control of the 3xP3/Hsp70 promoter (3xP3) was the transformation marker. PGUY1 refers to the native Guy1 promoter (Criscione et al., 2013). Note that the only difference between nGuy1 and Guy1m is the point mutation in the first ATG. PbZip1 refers to a promoter derived from an An. stephensi gene (Genbank JQ266223) and this promoter is used to drive early zygotic expression of the transgene (Figure 3). The C-tag and N-tag refer to the eight residue Strep II tag (Lichty et al., 2005) placed at either the C- or N-terminus of the GUY1 protein. A stretch of eight glycine residues were placed between the Strep II tag and the GUY1 protein (Supplementary file 3). The number of transgenic males and females were total counts from screens performed on all lines of each construct (Supplementary files 1 and 2).