cDNA library construction of the sweetpotato whitefly, Bemisia tabaci, using Gateway System

Tuesday, November 12, 2013
Exhibit Hall 4 (Austin Convention Center)
Na-Yeon Ko , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Moon Nam , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Sang-Eun Park , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Tae-Hee Ryu , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Jeong-Gon Kim , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Hyo-Seob Shin , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Hye-Ri Kwon , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Mi-Ja Seo , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Hyoun-Sub Lim , 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
Sweet potato whitefly (Bemisia tabaci) is one of the major and alien insect pests acting as viral vector in Korea. B. tabaci can be transmitted about 100 kinds of plant viruses including tomato yellow leaf curl virus (TYLCV) in tomato. In order to control for these B. tabaci, we mainly spray agricultural chemicals. However, the chemical control of B. tabaci is becoming problem because whitefly rapidly develops resistance expression to agricultural chemicals. A lot of insect pest control methods using RNA interference (RNAi)-based technology has been studied in recent years. Besides, As well as, RNAi technology can be helpful to study the target genes with applying several methods such as, injection of dsRNA into insect body, dsRNA feeding and transgenic plant. In this study we attempted cDNA library construction using gateway system for the using RNAi. When we use oligo d(T) primer, we can confirmed the titer of 1.4⨉104 colony/ml and insert sizes with 1.10kb and 1.27kb. We currently tried to little modify the method that is constructed a cDNA library with smaller size of insert cDNA using attB-N25 random primer and sonication. Subsequently, these cDNA libraries may be helpful to study target gene with VIGS (virus-induced gene silencing) using reverse genetics. Also, the most effective gene selection among them may be used for insect pest control.
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