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Theoretical and Applied Genetics

, Volume 117, Issue 1, pp 75–88 | Cite as

Expression of a Bacillus thuringiensis toxin (cry1Ab) gene in cabbage (Brassica oleracea L. var. capitata L.) chloroplasts confers high insecticidal efficacy against Plutella xylostella

  • Cheng-Wei LiuEmail author
  • Chin-Chung Lin
  • Jinn-Chin Yiu
  • Jeremy J. W. Chen
  • Menq-Jiau TsengEmail author
Original Paper

Abstract

Chloroplast genetic engineering is an environmentally friendly approach, where the foreign integrated gene is often expressed at a higher level than nuclear transformation. The cry1Ab gene was successfully transferred into the cabbage chloroplast genome in this study. The aadA and cry1Ab genes were inserted into the pASCC201 vector and driven by the prrn promoter. The cabbage-specific plastid vectors were transferred into the chloroplasts of cabbage via particle gun mediated transformation. Regenerated plantlets were selected by their resistance to spectinomycin and streptomycin. According to antibiotic selection, the regeneration percentage of the two cabbage cultivars was 4–5%. The results of PCR, Southern, Northern hybridization and western analyses indicated that the aadA and cry1Ab genes were not only successfully integrated into the chloroplast genome, but functionally expressed at the mRNA and protein level. Expression of Cry1Ab protein was detected in the range of 4.8–11.1% of total soluble protein in transgenic mature leaves of the two species. Insecticidal effects on Plutella xylostella were also demonstrated in cry1Ab transformed cabbage. The objectives of this study were to establish a gene transformation system for Brassica chloroplasts, and to study the possibility for insect-resistance in dicot vegetables using chloroplast gene transformation.

Keywords

Transgenic Plant Chloroplast Genome Spectinomycin Plastid Transformation Transplastomic Plant 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We gratefully acknowledge the support provided by Professor Liang-Jwu Chen for the donation of plasmid pKcBn. This research was supported by funds from the National Science Council, Taiwan (NSC 94-2752-B-005-005-PAE to Menq-Jiau Tseng).

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Department of Post-Modern AgricultureMing Dao UniversityChang HuaTaiwan ROC
  2. 2.Institute of Molecular BiologyNational Chung-Hsing UniversityTaichungTaiwan ROC
  3. 3.Department of HorticultureNational Ilan UniversityI-LanTaiwan ROC
  4. 4.Institute of Biomedical SciencesNational Chung-Hsing UniversityTaichungTaiwan ROC
  5. 5.Department of HorticultureNational Chung-Hsing UniversityTaichungTaiwan ROC

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