Expression of Cry1Ab and Cry2Ab by a polycistronic transgene with a self-cleavage peptide in rice

PLoS One. 2014 Oct 15;9(10):e110006. doi: 10.1371/journal.pone.0110006. eCollection 2014.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus thuringiensis / genetics*
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Endotoxins / genetics*
  • Endotoxins / metabolism
  • Hemolysin Proteins / genetics*
  • Hemolysin Proteins / metabolism
  • Insecticide Resistance / genetics*
  • Moths / genetics
  • Oryza / genetics
  • Pest Control, Biological / methods*

Substances

  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Endotoxins
  • Hemolysin Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis

Grants and funding

This research was supported by The National Natural Science Foundation of China (31021003, http://www.nsfc.gov.cn/) received by Zc Shen. The funders had no role in study design,data collection and analysis, decision to publish, or preparation of the manuscript.