Synthesis of an HIV-1 Tat transduction domain-rotavirus enterotoxin fusion protein in transgenic potato.
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.
Kim
T-G
TG
Department of Biochemistry and Microbiology and Center for Molecular Biology and Gene Therapy, School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA.
Langridge
W H R
WH
D001483:Base Sequence; D017931:DNA Primers; D004768:Enterotoxins / Q000235:genetics; D015696:Gene Products, tat / Q000096:biosynthesis*; D015497:HIV-1 / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis; D015183:Restriction Mapping; D012401:Rotavirus / Q000235:genetics; D011198:Solanum tuberosum / Q000235:genetics* / Q000821:virology; D014170:Transformation, Genetic; D054322:tat Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D004768:Enterotoxins; D015696:Gene Products, tat; D011993:Recombinant Fusion Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus