Expression of fluorescent proteins in homozygous <i>A. ludens</i> TTSS and SSIS.

<p>Images of the same individual flies from indicated lines under brightfield (BF; A-E), and epifluorescent microscopy using the Texas Red (TXR) filter for DsRed fluorescence (F-J), and the YFP filter for EGFP fluorescence (K-O). See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109690#s2" target="_blank">Materials and Methods</a> for filter specifications.</p

Comparison of population growth rates.

<p>Relative difference of λ between TTSS, SSIS, and WT strains is depicted.</p

Site-specific integration in <i>A. ludens</i> mediated by <i>phiC31</i> integrase.

<p>A) Schematic of the <i>pXLBacII[attP-PUbDsRed]</i> target site vector genomic integration recombining with the <i>pSL_3pB-attB-PUbEGFP</i> donor vector, at their respective <i>attP</i> and <i>attB</i> sites, in the presence of <i>phs-dphiC31</i> integrase helper plasmid, resulting in recombinant <i>attR</i> and <i>attL</i> sites within the SSIS expressing DsRed and EGFP. B) PCR amplification products using the JS01_F and JS01_R <i>attL</i> site primers in the <i>attP</i>_2-M6y and <i>attP</i>_F7 TTSS strains and the <i>int</i>_2-M6y and <i>int</i>_F7 SSIS strains. PCR products are observed only in the SSIS lines (bottom left), having sequences corresponding to the <i>attL</i> site with the core TTG in bold (bottom right).</p

Expression of DsRed fluorescent protein in pupae.

<p>Two day old pupae of the <i>A. ludens</i> Y-linked TTSS <i>attP</i>_2-M6y (A and B) and the TTSS <i>attP_</i>F7 (C and D) are shown under brightfield (A, C) and epifluorescence optics using the TXR filter (B, D).</p

Demographic analysis.

<p>A) Diagram of the <i>A. ludens</i> stage-structured life cycle with five transition stages (arrows between nodes). P<i>_i</i> represents the proportion of individuals in stage <i>i</i> that move to stage <i>i</i>+1 from time <i>t</i> to time <i>t</i>+1 (P₁  =  transition frequency from eggs to neonate larvae; P₂  =  from L1 to L3; P₃  =  from L3 to pupae; P₄  =  from pupae to adult), S₅ is the proportion of individuals that remain in stage <i>i</i> from time <i>t</i> to time <i>t</i>+1, and F₅ is the number of eggs produced per individual in stage <i>i</i> from time <i>t</i> to time <i>t</i>+1 (generational overlap); B) Stage-structured population transition matrix model. The population-projection matrix (A) is multiplied by the stage-distribution vector (N_t) of the five elements, which represents the number of individuals in each phase at time <i>t</i>, resulting in the estimation of λ; C) Relative difference of population growth rates λ between transgenic target-site strain (TTSS) and WT strains.</p

Vector insertion site sequencing of transgenic target site strains.

<p>Vector insertion site sequencing of transgenic target site strains.</p