EGFP expression at the trochophore stage reflects endogenous <i>tuba</i> expression in tuba::egfp^vbci1 animals.

Abstract

<p>(<b>A</b>) Scheme of donor vector pMos{tuba::egfp}^frkt707 containing 4.4 kB of DNA upstream of the <i>alpha-tubulin (tuba)</i> start codon (compare to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093076#pone-0093076-g002" target="_blank">Figure 2A</a>). (<b>B, C</b>) Expression of endogenous <i>tuba</i> in the prototroch (pt; arrows), apical ciliary tuft (at) and paratrochs (pat; arrowheads) of <i>Platynereis</i> larvae at the trochophore (B) and metatrochophore (C) stages. (<b>D, E</b>) Expression of <i>egfp</i> RNA (D) and EGFP protein in tuba::egfp^vbci1 trochophore larvae perfectly reflects endogenous <i>tuba</i> expression. (<b>F</b>) <i>egfp</i> expression in tuba::egfp^vbci1 metatrochophore larvae remains restricted to the prototroch (arrow); (<b>G</b>) Stable inheritance of the construct, as evidenced by Southern blots of carriers from three different generations, probed with a labeled <i>egfp</i> fragment.</p