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Seminested PCR for the detection and analysis of flue-cured tobacco (Nicotiana tabacum) expressing a chitinase transgene

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Abstract

Standard PCR was ineffective in detecting a baculovirus-derived chitinase transgene in the T1 generation of chitinase-expressingNicotiana tabacum cv. CF80 after leaves were flue-cured at high temperatures. Consequently, a seminested PCR method was developed using fresh leaves from T2 generation plants also expressing the chitinase protein. Seminested PCR was highly effective in detecting the chitinase transgene in fresh leaves ofN. tabacum cvs. Xanthi-nc and K326 and in both fresh and flue-cured leaves ofN. tabacum cv. CF80.

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Authors and Affiliations

  1. Agricultural College of Henan Agricultural University, Wenhua Road No. 95, 450002, Zhengzhou City Henan Province, China

    Jiao Shi & Jin Feng Han

  2. NERC Centre for Ecology and Hydrology, Mansfield Road, OX1 3SR, Oxford, UK

    Martin Naylor, Mary-Lou Edwards & Ian Cooper

Authors
  1. Jiao Shi
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  2. Martin Naylor
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  3. Mary-Lou Edwards
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  4. Jin Feng Han
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  5. Ian Cooper
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Corresponding author

Correspondence to Mary-Lou Edwards.

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Shi, J., Naylor, M., Edwards, ML. et al. Seminested PCR for the detection and analysis of flue-cured tobacco (Nicotiana tabacum) expressing a chitinase transgene. Plant Mol Biol Rep 20, 419 (2002). https://doi.org/10.1007/BF02772131

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  • DOI: https://doi.org/10.1007/BF02772131

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