Figure 1
From: RNA activation in ticks
Full-length ORF of HlemCHT cDNA. (a) Strategy for obtaining full-length ORF of HlemCHT. (1) Predicted exons and introns on the upstream sequence of the putative endochitinase-like gene (S03044-17L21 identified in the cDNA library, namely, HlemCHT) were retrieved from Chromosome 3 (Chr #3) after a nucleotide BLAST search of a partial ORF sequence against both local and NCBI nr databases of Haemaphysalis longicornis genome. (2) Forward and reverse primers (Red arrows) were designed to target sequences before the predicted starting methionine, and sequences at about 150 bp downstream from the end of the partial ORF fragment respectively. (3) Amplified cDNA was sequenced after cloning into a vector. (4) Assembly of newly sequenced 5ʹ-cDNA fragment and already existing cDNA fragment into a contig sequence was done using CAP3 Sequence Assembly Program to obtain the full-length ORF. (b) HlemCHT gene structure showing the positions the RNAi target site (Green) within the full-length ORF (Orange) and the RNAa target site (Yellow) at the 3′-UTR. Also, within the ORF is a conserved sequence (Blue) in other ixodid tick species’ endochitinase-like genes.
