Production of HIV-1 p24 protein in transgenic tobacco plants.
The production of antigens for vaccines in plants has the potential as a safe and cost-effective alternative to traditional production systems. Toward the development of a plant-based expression system for the production of human immunodeficiency virus type I (HIV-1) p24 capsid protein, the p24 gene was introduced into the genome of tobacco plants using Agrobacterium tumefaciens-mediated gene transfer. Southern blot analyses confirmed the presence of the p24 coding sequence within the genome of transgenic lines. Western blot analysis of protein extracts from transgenic plants identified plant-expressed p24 protein that cross-reacted with a p24-specific monoclonal antibody, thus confirming the maintenance of antigenicity. Quantification of the p24 protein using enzyme-linked immunosorbent assay (ELISA) estimated yields of approx 3.5 mg per g of soluble leaf protein. Similar accumulation levels of p24 were also detected in T1 plants, confirming that the p24 gene is transmitted stably. Our results indicate that plant-based transgenic expression represents a viable means of producing p24 for the development of HIV vaccine and for use in HIV diagnostic procedures.
Zhang
G Gary
GG
Department of Biology, York University, Toronto, Ontario, Canada. g.zhang@genesis.co.nz
Rodrigues
Lauren
L
Rovinski
Benjamin
B
White
K Andrew
KA
D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000821:virology; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D018744:DNA, Plant / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D015870:Gene Expression; D005822:Genetic Vectors; D018745:Genome, Plant; D016655:HIV Core Protein p24 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism* / Q000821:virology
D018744:DNA, Plant; D016655:HIV Core Protein p24; D011993:Recombinant Fusion Proteins