Title

Stable expression of Gal/GalNAc lectin of Entamoeba histolytica in transgenic chloroplasts and immunogenicity in mice towards vaccine development for amoebiasis

Authors

Authors

S. Chebolu;H. Daniell

Comments

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Abbreviated Journal Title

Plant Biotechnol. J.

Keywords

amoebiasis vaccine; genetically modified crops; plant-made vaccines; tobacco; LACTUCA-SATIVA L.; TOBACCO CHLOROPLASTS; PLASTID TRANSFORMATION; PROTECTIVE ANTIGEN; ADHERENCE LECTIN; LIGHT SUBUNIT; GENE; PROTEIN; GENOME; PEPTIDE; Biotechnology & Applied Microbiology; Plant Sciences

Abstract

Chloroplast genetic engineering offers several advantages, including high levels of transgene expression, transgene containment via maternal inheritance and multigene engineering in a single transformation event. Entamoeba histolytica infects 50 million people, causing about 100 000 deaths annually, but there is no approved vaccine against this pathogen. LecA, a potential target for blocking amoebiasis, was expressed for the first time in transgenic plants. Stable transgene integration into chloroplast genomes and homoplasmy were confirmed by polymerase chain reaction and Southern blot analyses. LecA expression was evaluated by Western blots and quantified by enzyme-linked immunosorbent assay (up to 6.3% of total soluble protein or 2.3 mg LecA/g leaf tissue). Subcutaneous immunization of mice with crude extract of transgenic leaves resulted in higher immunoglobulin G titres (up to 1 : 10 000) than in previous reports. An average yield of 24 mg of LecA per plant should produce 29 million doses of vaccine antigen per acre of transgenic plants. Such high levels of expression and immunogenicity should facilitate the development of a less expensive amoebiasis vaccine.

Journal Title

Plant Biotechnology Journal

Volume

5

Issue/Number

2

Publication Date

1-1-2007

Document Type

Article

Language

English

First Page

230

Last Page

239

WOS Identifier

WOS:000244280100003

ISSN

1467-7644

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