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Improved expression of recombinant GFP using a replicating vector based on Beet curly top virus in leaf-disks and infiltrated Nicotiana benthamiana leaves

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Abstract

Recombinant green fluorescent protein (GFP) with a molecular mass of 29 kDa was transiently expressed in Agrobacterium-inoculated leaf-disks prepared from Nicotiana benthamiana plants. Expression of GFP from the Cauliflower mosaic virus (CaMV) 35 S promoter within a replicating vector based on the geminivirus Beet curly top virus (BCTV) was more than 3 times higher than from a control, non-replicating vector. Use of the Cassava vein mosaic virus (CsVMV) promoter in the BCTV replicating vector increased the expression of recombinant GFP 320% at the transcript level, compared to use of the control CaMV 35 S promoter. Expression of recombinant GFP from Agrobacterium-inoculated leaf-disks of N. benthamiana was further enhanced up to 240% in the presence of post-transcriptional gene silencing suppressor p19.

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Acknowledgements

This work was supported by grants from the Biogreen 21 project, from the BK21 project, and from the Korea Science and Engineering Foundation through the Plant Metabolism Research Center, Kyung Hee University.

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Authors and Affiliations

  1. Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University, Suwon, 449-701, Korea

    Kyung Il Kim & In Sik Chung

  2. Department of Biology, University of Texas at San Antonio, TX, 78249, USA

    Garry Sunter

  3. Department of Molecular Genetics and Plant Biotechnology Center, Ohio State University, Columbus, OH, 43210, USA

    David M. Bisaro

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  1. Kyung Il Kim
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  2. Garry Sunter
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  3. David M. Bisaro
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  4. In Sik Chung
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Correspondence to In Sik Chung.

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Kim, K.I., Sunter, G., Bisaro, D.M. et al. Improved expression of recombinant GFP using a replicating vector based on Beet curly top virus in leaf-disks and infiltrated Nicotiana benthamiana leaves. Plant Mol Biol 64, 103–112 (2007). https://doi.org/10.1007/s11103-007-9137-z

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  • DOI: https://doi.org/10.1007/s11103-007-9137-z

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