The analysis of mitochondrial genomes may result in significant insights for both organismal and molecular evolution. The development of PCR primers to amplify full-length mitochondrial genomes has significant implications for various studies performed with invertebrates. Due to the difficulties of isolating sufficient purified mtDNA from small invertebrates, mitochondrial complete genomes have not been explored as fully in invertebrates as in vertebrates. The aim of this study is the characterization of the structure and evolution of 16S sequences of the horn fly mtDNA and the development of suitable primers for recovering the complete mitochondrial genome of Haematobia irritans and related species. For this purpose, partial 16S gene from Haematobia was amplified using the insect mtDNA universal primers 16Sar and ND1A. The amplification yielded a 814pb fragment which was sequenced by direct sequencing of the PCR product. Partial sequence of the ND1 gene, complete tRNALeu(UAG) gene and partial 16S sequence were analyzed. Comparative analysis among Haematobia and homologous sequences available in GenBank for other Calyptratae species were performed in order to identify conserved regions for designing reliable primers for the optimization of the Long-PCR technique. We expect that the optimization of Long-PCR amplification of Haematobia irritans mtDNA would provide a reliable strategy for complete sequencing of this genome. This approach could also be useful for the analysis of molecular evolution and population genetics based on mtDNA markers for other closely related species, including other parasitic lineages from Muscidae. Financial support: CNPq/PROFIX;FAPESP.
Species 1: Diptera Muscidae Haematobia irritans (horn fly)
Keywords: Long-PCR, mitochondrial DNA
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