title: Expression of Cry1Ab and Cry2Ab by a polycistronic transgene with a self-cleavage peptide in rice.
abstract: Insect resistance to Bacillus thuringiensis (Bt) crystal protein is a major threat to the long-term use of transgenic Bt crops. Gene stacking is a readily deployable strategy to delay the development of insect resistance while it may also broaden insecticidal spectrum. Here, we report the creation of transgenic rice expressing discrete Cry1Ab and Cry2Ab simultaneously from a single expression cassette using 2A self-cleaving peptides, which are autonomous elements from virus guiding the polycistronic viral gene expression in eukaryotes. The synthetic coding sequences of Cry1Ab and Cry2Ab, linked by the coding sequence of a 2A peptide from either foot and mouth disease virus or porcine teschovirus-1, regardless of order, were all expressed as discrete Cry1Ab and Cry2Ab at high levels in the transgenic rice. Insect bioassays demonstrated that the transgenic plants were highly resistant to lepidopteran pests. This study suggested that 2A peptide can be utilized to express multiple Bt genes at high levels in transgenic crops.
authors.affiliation: State Key Laboratory of Rice Biology, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Qichao
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Rice Biology, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Minghong
authors.initials: M
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Rice Biology, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Tan
authors.affiliation: College of Life Science, Shanxi Agricultural University, Taigu, China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Rice Biology, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: Shen
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism; D006460:Hemolysin Proteins / Q000235:genetics* / Q000378:metabolism; D007305:Insecticide Resistance / Q000235:genetics*; D009036:Moths / Q000235:genetics; D012275:Oryza / Q000235:genetics; D010572:Pest Control, Biological / Q000379:methods*
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis