꿀벌에 대한 dsRNA의 급성섭식독성 평가

• Published in: Hanguk hwangyeong nonghak hoeji Vol. 36; no. 4; pp. 241 - 248
• Main Authors: 임혜송, Hye Song Lim, 정영준, Young Jun Jung, 김일룡, Il Ryong Kim, 김진, Jin Kim, 유성민, Sungmin Ryu, 김반니, Banni Kim, 이중로, Jung Ro Lee, 최원균, Wonkyun Choi
• Format: Journal Article
• Language: Korean
• Published: 한국환경농학회 31.12.2017
• Subjects: Acute oral toxicity; Apis mellifera; dsRNA; Living modified organisms; 농학; Apis mellifera; dsRNA; Living modified organisms; Acute oral toxicity
• ISSN: 1225-3537; 2233-4173
• DOI: 10.5338/KJEA.2017.36.4.36

BACKGROUND: RNA interference (RNAi) eliminates or decreases gene expression by disrupting the target mRNA or by interfering with translation. Recently, RNAi technique was applied to generate new crop traits which provide protection against pests. To establish the environmental risk assessment protocol of RNAi LMO in lab scale, we developed dsRNA expression system using E. coli and tested acute oral toxicity assay to honey. METHOD AND RESULTS: The dsRNA expression vector, L4440, was chosen and cloned 240 bp of Snf7 and GFP gene fragment. To develop the maximum dsRNA induction condition in E. coli, we tested induction time, temperature and IPTG concentration in media. To estimate the risk assessment of dsRNA to honey bee, it has been selected and cultured with dsRNA supplement for 48 hours according to OECD guideline. As a result, the optimum condition of dsRNA induction was 37℃, 4 hours and 0.4 mM IPTG concentration and the difference between Snf7 and GFP dsRNA molecules from E. coli was not significant in survival and behavior to honey bee. Furthermore, blast search results indicated that effective match of predicted dsRNA fragments were not existed in honey bee genome. CONCLUSION: In this study, we developed and tested the acute oral toxicity of dsRNA using E. coli expression system to honey bee.