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Construction of modified Bacillus thuringiensis cry1AC genes based on cry1-5 genes through multi site-directed mutagenesis

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  • URLhttps://db.koreascholar.com/Article/Detail/291445
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한국응용곤충학회 (Korean Society Of Applied Entomology)
초록

Bt crystal proteins, encoded by cry genes, are a group of insecticidal proteins unique in the Gram-positive and spore-forming bacterium, Bacillus thuringiensis. These cry genes are widely applied as one of the most successful candidates for constructing transgenic plants resistant to pest insects. In our previous report, we found Cry1-5 had high insecticidal activity against Spodoptera larvae although its amino acid sequences showed high similarity (97.9%) to those of Cry1Ab which had low activity. In comparison with Cry1Ac, Cry1-5 had 12 different residues in domain Ⅰ and domain Ⅱ, and we focused on domain Ⅰand domain Ⅱ regions and designed 10 mutagenic primers to change 12 residues. Through multi site-directed mutagenesis, we mutated the modified cry1Ac gene by plant codon usage in pOBⅠ-Mod-cry1Ac based on cry1-5 and constructed 63 various mutant cry genes. In the further study, we will express those mutant proteins as a fusion form with polyhedrin using baculovirus expression system and subsequently do bioassay to Spodoptera larvae.

저자
  • Hong Guang Xu(Department of Agricultural Biotechnology, Seoul National University)
  • Jong Yul Roh(Department of Agricultural Biotechnology, Seoul National University)
  • Jae Young Choi(Research Institute for Agriculture and Life Sciences, Seoul National University)
  • Hee Jin Shim(Department of Agricultural Biotechnology, Seoul National University)
  • Yong Wang(Department of Agricultural Biotechnology, Seoul National University)
  • Qin Liu(Department of Agricultural Biotechnology, Seoul National University)
  • Byung Rae Jin(College of Natural Resources and Life Science, Dong-A University)
  • Yeon Ho Je(Department of Agricultural Biotechnology, Seoul National University)