cDNA library construction of the Asian ladybird beetle using gateway cloning system

Tuesday, November 17, 2015
Exhibit Hall BC (Convention Center)
Yu-Bin Jung , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Jung Kyu Kim , Chungnam National University, Daejeon, South Korea
Chan-yeong Kang , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Jeong Hee Kim , Chungnam National University, Daejeon, South Korea
Ji Hyun Min , Chungnam National University, Daejeon, South Korea
Il Hyun Byun , Chungnam National University, Daejeon, South Korea
Hyun Ju Jang , Chungnam National University, Daejeon, South Korea
Min Gyu Cho , Chungnam National University, Daejeon, South Korea
Tae-Hee Ryu , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Yong-Man Yu , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Young-Nam Youn , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
The Asian ladybird beetle (Harmonia axyridis, Coccinellidae, Coleoptera) has been well known for biological control agents against many aphid species in the greenhouses as well as in the fields. However, the gene function of H. axyridis has not been commonly researched. So, we tried to know about gene functions of H. axyridis using RNA interference (RNAi) technique. It is supposed that RNAi technology can be helpful to study on the gene functional analysis. In this study, we produce cDNA library construction using gateway cloning system for selecting target gene in order to know about gene function in H. axyridis for using RNAi. As a result, the 200bp of insert size, which is appropriate for RNAi was confirmed. Consequently, H. axyridis of cDNA library with the titer of 2.58 x 106 clones were completed. And the random cDNA is cloning to transcription vector (LITMUS 28i vector) for in vitro transcription. And it was injected dsRNA to 4th larvae of H. axyridis by using 5μl micro syringe and observed the phenotype appears at the adult stage. It is supposed that this methods may be useful for searching for the gene functional analysis in H. axyridis as well as other insects.

Keywords: Harmonia axyridis, RNA interference, cDNA library

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