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Synthesis of an HIV-1 Tat transduction domain-rotavirus enterotoxin fusion protein in transgenic potato

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

A DNA fragment encoding a 12-amino acid (aa) HIV-1 Tat transduction peptide fused to a 90-aa murine rotavirus NSP4 enterotoxin protein (Tat-NSP490) was transferred to Solanum tuberosum by Agrobacterium tumefaciens-mediated transformation. The fusion gene was detected in the genomic DNA of transformed plant leaf tissues by PCR DNA amplification. The Tat-NSP490 fusion protein was identified in transformed tuber extracts by immunoblot analysis using anti-NSP490 and anti-Tat as the primary antibodies. Enzyme-linked immunosorbent assay results showed that the Tat-NSP490 fusion protein made up to 0.0015% of the total soluble tuber protein. The synthesis of Tat-NSP490 fusion protein in transformed potato tuber tissues demonstrates the feasibility of plant cell delivery of the HIV-1 Tat transduction domain as a carrier for non-specific targeting of fused antigens to the mucosal immune system.

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Abbreviations

APC :

Antigen-presenting cells

BA :

Benzyladenine

BSA :

Bovine serum albumin

CT :

Cholera toxin

CTB :

Cholera toxin B subunit

CTL :

Cytotoxic T lymphocytes

2,4-D :

2,4-Dichlorophenoxyacetic acid

ELISA :

Enzyme-linked immunosorbent assay

HIV-1 :

Human immunodeficiency virus type 1

MHC :

Major histocompatibility complex

IAA :

Indole-3-acetic acid

NAA :

α-Naphthaleneacetic acid

NPT II :

Neomycin phosphotransferase II

NSP4 :

Rotavirus enterotoxin non-structural protein

PBS :

Phosphate-buffered saline

PBST :

Phosphate-buffered saline containing 0.05% Tween-20

PTD :

Protein transduction domain

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Acknowledgements

The authors would like to thank Dr. Stephen Dowdy, Howard Hughes Medical Institute, University of California, San Diego for contribution of the plasmid pHA-Tat and Dr. Mary Estes, Baylor College of Medicine, for providing plasmid pCR2.1-NSP4 containing simian virus SA 11 gene 10 encoding the full-length NSP4175 protein. This project was supported by a National Medical Testbed-subcontract to W.H.R.L. The views, opinions and/or findings contained in this report are those of the authors and should not be construed as a position, policy, decision or endorsement of the National Medical Technology Tested INC.

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Authors and Affiliations

  1. Department of Biochemistry and Microbiology and Center for Molecular Biology and Gene Therapy, School of Medicine, Loma Linda University, Loma Linda, CA, 92350, USA

    T.-G. Kim & W. H. R. Langridge

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  1. T.-G. Kim
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  2. W. H. R. Langridge
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Correspondence to W. H. R. Langridge.

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Communicated by W.A. Parrott

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Kim, TG., Langridge, W.H.R. Synthesis of an HIV-1 Tat transduction domain-rotavirus enterotoxin fusion protein in transgenic potato. Plant Cell Rep 22, 382–387 (2004). https://doi.org/10.1007/s00299-003-0697-3

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  • DOI: https://doi.org/10.1007/s00299-003-0697-3

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