Plasmid DNA associated with crystal protein coding in Bacillus thuringiensis CAB565 and CAB566

Tuesday, November 17, 2015
Exhibit Hall BC (Convention Center)
You-Kyoung Lee , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Na-Young Jin , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Jun-Hack Jeon , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Yu-Seop Kim , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Bo-Ram Lee , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Hee Ji Kim , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Soo Jeung Ahn , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Young-Nam Youn , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Yong-Man Yu , Department of Applied Biology, Chungnam National University, Daejeon, South Korea
Bacillus thuringiensis is widely known that it is the gram-positive bacteria to produce a parasporal crystal protein comprised the endotoxin in spore and the most effective bio-insecticide. Cry gene involved in the production of crystal protein is mainly presented in the large size of the plasmid DNA. We were examined the plasmid DNA associated with crystal protein coding of B. t. subsp. kurstaki CAB565 and B. t. subsp. aizawai CAB566. CAB565 and CAB566 strains have only cry1 gene indicating the insecticidal activity to lepidopteran. As a result of quantification by real-time PCR analysis in plasmid DNA of the two strains, CAB565 strain has cry1Aa, cry1Ac and cry1I genes, and CAB565 strain has cry1Aa, cry1C and cry1D genes. By using agarose gel electrophoresis after real-time PCR, we confirmed that two strains having gene was expressed more strongly. In order to extract the cry gene located in plasmid DNA, 23.1kbp and higher size bands of the two strains were extracted from the gel. As a result of agarose gel electrophoresis, it was confirmed that two strains showing plasmid DNA bands and presentation of cry genes in plasmid DNA were confirmed using each of the primer by a PCR.

Key Words : B. thuringiensis subsp. kurstaki, B. thuringiensis subsp. aizawai, plasmid DNA

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