Effect of codon optimization and subcellular targeting on Toxoplasma gondii antigen SAG1 expression in tobacco leaves to use in subcutaneous and oral immunization in mice.

BMC Biotechnology
Melina Laguía-BecherMarina Clemente

Abstract

Codon optimization and subcellular targeting were studied with the aim to increase the expression levels of the SAG178-322 antigen of Toxoplasma gondii in tobacco leaves. The expression of the tobacco-optimized and native versions of the SAG1 gene was explored by transient expression from the Agrobacterium tumefaciens binary expression vector, which allows targeting the recombinant protein to the endoplasmic reticulum (ER) and the apoplast. Finally, mice were subcutaneously and orally immunized with leaf extracts-SAG1 and the strategy of prime boost with rSAG1 expressed in Escherichia coli was used to optimize the oral immunization with leaf extracts-SAG1. Leaves agroinfiltrated with an unmodified SAG1 gene accumulated 5- to 10-fold more than leaves agroinfiltrated with a codon-optimized SAG1 gene. ER localization allowed the accumulation of higher levels of native SAG1. However, no significant differences were observed between the mRNA accumulations of the different versions of SAG1. Subcutaneous immunization with leaf extracts-SAG1 (SAG1) protected mice against an oral challenge with a non-lethal cyst dose, and this effect could be associated with the secretion of significant levels of IFN-gamma. The protection was increased ...Continue Reading

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Citations

Jun 28, 2011·PloS One·Mariana G CoriglianoMarina Clemente
Aug 21, 2013·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Abdullah MakhzoumJocelyne Trémouillaux-Guiller
Aug 23, 2012·Journal of Biomedicine & Biotechnology·Marina Clemente, Mariana G Corigliano
Oct 8, 2013·Expert Review of Vaccines·Nian-Zhang ZhangXing-Quan Zhu
Oct 2, 2012·Plant Biotechnology Journal·María Del L YáconoMarina Clemente
Jun 5, 2015·Critical Reviews in Biotechnology·Khaled MoustafaJocelyne Trémouillaux-Guiller
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Jan 21, 2020·Comparative Immunology, Microbiology and Infectious Diseases·Abdol Sattar PaghehAhmad Daryani

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Methods Mentioned

BETA
PCR
transgenic
ELISA
enzyme-linked immunosorbent assay

Software Mentioned

Prism
Pro
- Pro analyzer
Pro analyzer
Gel
CAI Calculator

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