Mapping of Himar1 transposon insertion site. A. Location of primer pairs (AB1553-AB1554) and (AB1561-AB1562) designed to target the omp6 and omp10 genes respectively, in wild-type (WT) and transformed A. marginale. Based on sequencing results the Himar1 sequences are integrated in the chromosome of transformed A. marginale at nucleotide 245 after the first base of the omp10 start codon (arrows) and mCherry and aadA (Str/Spc resistant) genes are in the opposite orientation to omp10. B. Agarose Gel electrophoresis. gDNA isolated from ISE6 tick cells infected with wild-type (WT) and transformed A. marginale, was used as template for PCR amplification with primers shown in A. (Lane 1) 100 bp/1Kb DNA ladder, omp6 amplicons in transformed (lane 2) and WT (lane3) A. marginale were of the same size 492 bp. The omp10 amplicon in transformed A. marginale (lane 4) was 2805 bp, while in wild type was 969 bp (lane 5).