HIV-1 gp120 V3 cholera toxin B subunit fusion gene expression in transgenic potato

Tae Geum Kim; Andreas Gruber; William H.R. Langridge

doi:10.1016/j.pep.2004.04.014

HIV-1 gp120 V3 cholera toxin B subunit fusion gene expression in transgenic potato

Tae Geum Kim, Andreas Gruber, William H.R. Langridge

Biochemistry Division

Research output: Contribution to journal › Article › peer-review

Abstract

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 G_M1-ganglioside enzyme-linked immunosorbent assay (G_M1-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.

Original language	English
Pages (from-to)	196-202
Number of pages	7
Journal	Protein Expression and Purification
Volume	37
Issue number	1
DOIs	https://doi.org/10.1016/j.pep.2004.04.014
State	Published - Sep 2004

ASJC Scopus Subject Areas

Biotechnology

Keywords

Edible vaccine
HIV-1
Solanum tuberosum
gp120 V3 loop

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10.1016/j.pep.2004.04.014

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title = "HIV-1 gp120 V3 cholera toxin B subunit fusion gene expression in transgenic potato",

abstract = "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.",

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note = "Funding Information: The authors thank Dr. John Mekalanos, Harvard University Medical School, for the gift of plasmid pRT42 containing ctxAB from the Classical Vibrio cholerae strain 569B. The research reported in the manuscript was supported by an intramural grant from Loma Linda University to W.H.R.L.",

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HIV-1 gp120 V3 cholera toxin B subunit fusion gene expression in transgenic potato

Abstract

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Keywords

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