Selective whole genome amplification for mitogenome analysis of disease vectors

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K21358
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 42:Veterinary medical science, animal science, and related fields
• Research Institution: Hokkaido University
• Principal Investigator: Nakao Ryo 北海道大学, 獣医学研究院, 准教授 (50633955)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Keywords: 選択的ゲノム増幅法 / 節足動物 / マダニ / ミトゲノム / SWGA

Outline of Final Research Achievements

The purpose of this study was to develop a method to sequence complete mitochondrial genomes (mitogenomes) of ticks by using a selective whole genome amplification (SWGA) method. The tick mitogenome-specific primers were selected based on the alignment of tick mitogenome sequences available in the public database. The tick DNA was incubated with specific primers and phi29 DNA polymerase and the resulting products were sequenced on a MiSeq platform. The results indicated that SWGA could increase the relative abundance of mitochondrial DNA by more than 10,000 holds in comparison to chromosomal DNA. The developed method enables us to sequence complete tick mitogenomes in an easy and low-cost way. We proved that the method is applicable to many tick species, of which mitogenome sequences have not yet been reported.

Free Research Field

寄生虫学

Academic Significance and Societal Importance of the Research Achievements

開発した選択的ミトゲノム増幅技術により、簡便かつ安価にマダニのミトゲノム情報を取得することができることとなった。このことにより、世界中でマダニのミトゲノムの解読が進み、マダニの系統進化、集団遺伝構造を理解するための遺伝子情報の集積が期待できる。また、得られたゲノム情報を活用することで、精度の高いマダニの遺伝子同定技術の開発が可能となり、マダニが媒介する感染症の対策に貢献できる。また、本研究で確立した手法は、他の生物群にも応用可能であり、生物を対象とした広い研究分野への波及効果が期待できる。