Creation and characterization of <i>DmMterf3</i> knockout larvae.
2013-01-03T01:13:13Z (GMT) by
<p>(A) The <i>DmMterf3</i> locus and generation of a <i>DmMterf3</i> null mutant. <i>DmMterf3</i> is located on the third chromosome at cytological position 77C6. The construct (ko <i>DmMterf3</i>) used for ends-out homologous recombination is indicated by grey boxes, coding sequences in exons are indicated by black boxes and non-coding sequences in exons by white boxes. The gap between grey boxes represents the genomic region replaced by attP and a <i>white</i> marker gene. The white marker gene was subsequently removed by crossing to <i>cre</i>-recombinase expressing flies. (B) PCR analysis of wild-type and knockout alleles for <i>DmMterf3</i>. (C) Body size comparison of <i>DmMterf3</i> knockout larvae 6 days after egg-laying (ael) showing reduced size. (D) QRT-PCR analysis of <i>DmMterf3</i> transcript levels in control (white and grey bars) and <i>DmMterf3</i> KO larvae (black bars) at 3 and 6 days ael. (E) Q-PCR analysis of mtDNA levels in larvae at 3 and 6 days ael. (F) QRT-PCR analysis of steady-state levels of mitochondrial mRNAs normalized to the nuclear ribosomal protein 49 transcript levels in larvae at 3 and 6 days ael. Error bars indicate mean ± SEM (<sup>*</sup>p<0.05; <sup>**</sup>p<0.01; <sup>***</sup>p<0.001; n = 5).</p>