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

• Published in: Plant cell reports Vol. 22; no. 6; pp. 382 - 387
• Main Authors: Kim, T-G, Langridge, W H R
• Format: Journal Article
• Language: English
• Published: Germany Springer Nature B.V 01.01.2004
• Subjects: Amino acids; Bacteria; Base Sequence; Deoxyribonucleic acid; DNA; DNA Primers; Enterotoxins - genetics; Enzymes; Gene Products, tat - biosynthesis; HIV-1 - genetics; Human immunodeficiency virus 1; Immune system; NSP4 protein; NSP4-90 protein; Plant tissues; Plants, Genetically Modified - genetics; Polymerase Chain Reaction; Potatoes; Proteins; Recombinant Fusion Proteins - analysis; Recombinant Fusion Proteins - biosynthesis; Restriction Mapping; Rotavirus - genetics; Solanum tuberosum; Solanum tuberosum - genetics; Solanum tuberosum - virology; tat Gene Products, Human Immunodeficiency Virus; Tat protein; Transformation, Genetic
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-003-0697-3

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-NSP4(90)) 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-NSP4(90 )fusion protein was identified in transformed tuber extracts by immunoblot analysis using anti-NSP4(90) and anti-Tat as the primary antibodies. Enzyme-linked immunosorbent assay results showed that the Tat-NSP4(90) fusion protein made up to 0.0015% of the total soluble tuber protein. The synthesis of Tat-NSP4(90) 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.