극동산 뱀장어(Anguilla japonica)의 친자확인을 위한 유전자 마커 개발
Development of Microsatellite Markers for Parentage Analysis in the Japanese Eel Anguilla japonica
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초록
The Japanese eel Anguilla japonica is a highly valued research object that is important for aquaculture in Asia, including the Republic of Korea. However, few studies have been conducted analyzing parentage using microsatellite markers derived from the Japanese eel. We acquired Japanese eel genome data using next generation sequencing technology, and constructed a draft genome comprising 1,087 Mbp. Using the Simple Sequence Repeat Identification Tool program, 444,724 microsatellites were identified. Of these, 1,842 microsatellites located in the 3' untranslated region, which are stably inherited, were finally selected. Ninety-six primers were selected to validate polymorphism at these microsatellites, and 9 primers were finally identified for multiplex analysis. Using multiplex polymerase chain reaction with three different fluorescence chemistries, we performed parentage analysis of an artificial Japanese eel population. CERVUS software was used to calculate the logarithm of the odds (LOD) scores and the confidence of the parentage assignments. The results presented here show that 83 out of 85 paternity cases were assigned at 95% confidence to a candidate father and mother with LOD scores ranging from 4.79 to 28.2. This study provided a microsatellite marker-based assay for parentage analysis of Japanese eels, which will be useful for selective breeding and genetic diversity studies.
Abstract
The Japanese eel Anguilla japonica is a highly valued research object that is important for aquaculture in Asia, including the Republic of Korea. However, few studies have been conducted analyzing parentage using microsatellite markers derived from the Japanese eel. We acquired Japanese eel genome data using next generation sequencing technology, and constructed a draft genome comprising 1,087 Mbp. Using the Simple Sequence Repeat Identification Tool program, 444,724 microsatellites were identified. Of these, 1,842 microsatellites located in the 3' untranslated region, which are stably inherited, were finally selected. Ninety-six primers were selected to validate polymorphism at these microsatellites, and 9 primers were finally identified for multiplex analysis. Using multiplex polymerase chain reaction with three different fluorescence chemistries, we performed parentage analysis of an artificial Japanese eel population. CERVUS software was used to calculate the logarithm of the odds (LOD) scores and the confidence of the parentage assignments. The results presented here show that 83 out of 85 paternity cases were assigned at 95% confidence to a candidate father and mother with LOD scores ranging from 4.79 to 28.2. This study provided a microsatellite marker-based assay for parentage analysis of Japanese eels, which will be useful for selective breeding and genetic diversity studies.
참고문헌 (24)
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