HIV-1 gp120 V3 cholera toxin B subunit fusion gene expression in transgenic potato.
A cDNA fragment encoding the V3 loop of human immunodeficiency virus type-1 (HIV-1) envelope glycoprotein gp120 was fused to the cholera toxin B subunit gene (CTB-gp120) and transferred into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation. The CTB-gp120 fusion gene was detected in genomic DNA from transformed potato leaves by PCR DNA amplification. Synthesis and assembly of the CTB-gp120 fusion protein into oligomeric structures of pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding of CTB-gp120 fusion protein pentamers to intestinal epithelial cell membrane glycolipid receptors was quantified by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA results indicated that CTB-gp120 fusion protein made up 0.002-0.004% of the total soluble tuber protein. Synthesis of CTB-gp120 monomers and their assembly into biologically active oligomers in transformed potato tuber tissues demonstrates for the first time the expression of HIV-1 gp120 in plants and emphasizes the feasibility of using edible plant-based vaccination for protection against HIV-1 infection.
Center for Molecular Biology and Gene Therapy, Department of Biochemistry and Microbiology, School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA.
Tae-Geum
TG
Kim
Andreas
A
Gruber
William H R
WH
Langridge
D002772:Cholera Toxin / Q000235:genetics / Q000378:metabolism; D005677:G(M1) Ganglioside / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010936:Plant Extracts / Q000737:chemistry; D030821:Plants, Genetically Modified; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011198:Solanum tuberosum / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D015699:HIV Envelope Protein gp120; D010936:Plant Extracts; D011993:Recombinant Fusion Proteins; D005677:G(M1) Ganglioside; D002772:Cholera Toxin