Sequencing And Analysis Of Sinopodisma Houshana And Calliptamus Italicus Mitochondrial Genomes

Mitochondrial DNA has been widely used to study the population genetic structure, phylogeography, and molecular phylogenetic at various taxonomic levels since 1970s. The great progress of sequencing techniques made the dreams of sequencing massive taxa come to true today. At present there are 745 species of mitochondrial genomes from metazoan animals had been sequenced, including 49 species of Insecta. Among these species, only two come from Orthoptera, Locusta migratoria and Gryllotalpa orientalis. Accordingly, the more complete mitochondrial genome sequences of Orthoptera species are desirable to be sequenced and analyzed.Two Catantopidae grasshoppers, Sinopodisma houshana and Calliptamus italicus, have been choice to sequence and assembly using the strategy of sub-PCR based on the long PCR technology and clone sequencing method. The sequences of gene and protein composition, synonymous codons usage bias, tRNA and rRNA secondary structure etc, of the two mitochondrial genome were analyzed in details. Finally, the phylogenetic relationship of taxa with complete mitochondrial genome from Hexapoda was reconstructed by concatenated amino-acid sequences of 12 proteins downloaded from GenBank. Some conclusions drawn from the study as follows.1. The strategy of sub-PCR based on the long PCR here used to have solved the difficulty of mitochondrial DNA purification problem, reduces the risk of unintended amplification of nuclear mitochondrion-like pseudogenes. A rapid and accurate approach to sequence the complete insect mitochondrial DNA was established based on the strategy of combining sub-PCR with the clone sequencing method to make it more ease and stabile.2. A set of new universal primers designed in this study can be used to amplify and sequence the mitochondrial genome from Orthoptera, and has been successfully used to species, Sinopodisma houshana and Calliptamus italicus.3. The mitogenome length of Sinopodisma houshana and Calliptamus italicus is 15 818 bp and 15 667 bp, respectively. The nucleotide composition shows a high bias towards A+T. Both genome contents consist of the 13 protein-coding genes, 22 tRNAs,...