Analysis of transcription factor <i>wtrM.</i>

A. Phylogenetic tree of Wolbachia transcriptional regulator (wtr) genes. An unrooted maximum likelihood phylogenetic tree, constructed using PHYLIP, of transcriptional regulator genes in the wPip and wMel genomes from amino acid alignments. Bootstrap percentages for 100 trees are shown for each node. B. Transcription of wtrM in Culex mosquitoes. Expression of wtrM was validated by RNA extraction and PCR on the cDNA using primers internal to the open reading frame of the gene. There is no transcription of wtrM detectable in control untransfected ovaries (a) or carcasses (b) from Pel. There is a high level of transcription of wtrM in ovaries from untransfected Mol mosquitoes (c) but not in carcasses (d). In Pel mosquitoes transfected with wtrM by injection into the thorax (e, f) there is no transcription detected in ovaries (e) but strong expression in carcasses (f). Transfection of wtrM into the abdomen of Pel mosquitoes produced no detectable transcription in ovaries (g) but transcription readily detected in carcasses (h). An untransfected whole female of Pel (i) and Mol (j) are also shown. C. CPIJ005623 upregulation in transfected Culex females. Transcription of the Culex gene CPIJ005623 in Pel females transfected with the wPipMol gene wtrM, compared to Pel females transfected with the same plasmid but containing no insert. Difference was significant at p<0.01 using a Wilcoxon rank sum test.