Expression of active human epidermal growth factor (hEGF) in tobacco plants by integrative and non-integrative systems

Wirth, Sonia; Calamante, Gabriela; Mentaberry, Alejandro; Bussmann, Leonardo E.; Lattanzi, Mariano L.; Barañao, Lino; Bravo-Almonacid, Fernando

doi:10.1023/b:molb.0000012329.74067.ca

Cited by 59 publications

(42 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our results demonstrate that oral immunization of the transgenic rice seeds expressing A1aB1b-Crp-1 and -2 protein resulted in the generation of systemic unresponsiveness with a reduction of allergen-specific Th2-mediated IgE responses and histamine release. It has been demonstrated that the direct production of short peptides such as T cell epitope peptides with lengths of 10-20 aa is difficult for most expression systems of eukaryotic and prokaryotic cells (37). Therefore, our efforts in this study were initially focused on the expression of Crp-1 and -2 T cell epitope peptides in transgenic rice seeds.…”

Section: Discussionmentioning

confidence: 99%

A rice-based edible vaccine expressing multiple T cell epitopes induces oral tolerance for inhibition of Th2-mediated IgE responses

Takagi

Hiroi

Yang

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

206

137

View full text Add to dashboard Cite

Peptide immunotherapy using multiple predominant allergen-specific T cell epitopes is a safe and promising strategy for the control of type I allergy. In this study, we developed transgenic rice plants expressing mouse dominant T cell epitope peptides of Cry j I and Cry j II allergens of Japanese cedar pollen as a fusion protein with the soybean seed storage protein glycinin. Under the control of the rice seed storage protein glutelin GluB-1 promoter, the fusion protein was specifically expressed and accumulated in seeds at a level of 0.5% of the total seed protein. Oral feeding to mice of transgenic rice seeds expressing the T cell epitope peptides of Cry j I and Cry j II before systemic challenge with total protein of cedar pollen inhibited the development of allergen-specific serum IgE and IgG antibody and CD4 ؉ T cell proliferative responses. The levels of allergen-specific CD4 ؉ T cell-derived allergy-associated T helper 2 cytokine production of IL-4, IL-5, and IL-13 and histamine release in serum were significantly decreased. Moreover, the development of pollen-induced clinical symptoms was inhibited in our experimental sneezing mouse model. These results indicate the potential of transgenic rice seeds in production and mucosal delivery of allergen-specific T cell epitope peptides for the induction of oral tolerance to pollen allergens.Japanese cedar pollinosis ͉ peptide immunotherapy ͉ seed-specific expression

show abstract

Section: Discussionmentioning

confidence: 99%

A rice-based edible vaccine expressing multiple T cell epitopes induces oral tolerance for inhibition of Th2-mediated IgE responses

Takagi

Hiroi

Yang

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

206

137

View full text Add to dashboard Cite

show abstract

“…Among subcellular targets, ER possesses a favourable environment for the stable accumulation of recombinant proteins due to its low protease activity and the presence of molecular chaperones [64,65]. Targeting the protein to the ER can be successfully employed through the addition of the ER retention signal, KDEL (Lys-Asp-Glu-Leu), to the C-terminus of the considered protein, and it can lead to a several-fold increase in accumulation levels compared to targeting to the apoplast or cytosol in tobacco and other plants [66,67]. Therefore, the KDEL tetrapeptide ER-retention signal was fused to the C-terminus of LFchimera (Figure 1(A)) to facilitate the retention of the peptide within the ER lumen and increase its accumulation in plant cells.…”

Section: Peptide Purification and Antimicrobial Activity Evaluationmentioning

confidence: 99%

Recombinant expression of LFchimera antimicrobial peptide in a plant-based expression system and its antimicrobial activity against clinical and phytopathogenic bacteria

Chahardoli

Fazeli

Niazi‎

et al. 2018

Biotechnology & Biotechnological Equipment

View full text Add to dashboard Cite

Antimicrobial peptides (AMPs) with their broad and selective antimicrobial activity and lowered risk of resistance development in microbial populations are promining as a new alternative candidate to conventional antibiotics. Plant-based expression systems can be employed as cost-effective and high scale-up capacity production hosts for recombinant expression of AMPs. LFchimera is a chimerical peptide containing LFcin and LFampin antimicrobial peptides of bovine lactoferrin, and it has stronger bactericidal activity. Here, the LFchimera sequence was codon-optimized for tobacco and fused with endoplasmic reticulum retention signals along with a CaMV 35S promoter and then transferred by agrobacterium-mediated transformation. The integration and expression of the transgene in tobacco plants were confirmed by polymerase chain reaction (PCR) and RT-PCR, respectively. Recombinant production of the peptide was confirmed by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE), and peptide functional activity was confirmed by the antibacterial evaluation of total protein extracts against various clinical and phytopathogenic bacteria. Finally, LFchimera peptide was partially purified using an affinity column, and the antibacterial activity was shown. Taken together, these results confirmed that tobacco can be utilized for the recombinant production of antimicrobial peptides such as LFchimera.

show abstract

“…Higo et al (1993) tried to express human epidermal growth factor (hEGF) in tobacco plants, but the recombinant protein harvested only accounted for 0.001% of the total soluble proteins. Wirth et al (2004) constructed two different vectors that caused the recombinant protein to be targeted into either the cytoplasm or the apoplast. Recombinant hEGF with biological activity was accumulated in the endoplasmic reticulum at 0.1% of the total soluble proteins.…”

Section: Introductionmentioning

confidence: 99%

Expression and characteristic of synthetic human epidermal growth factor (hEGF) in transgenic tobacco plants

Bai

Zeng

et al. 2007

Biotechnol Lett

View full text Add to dashboard Cite

To improve the accumulation of recombinant human epidermal growth factor (hEGF) in transgenic tobacco, a highly effective vector was constructed and transformed via Agrobacterium tumefaciens. The hEGF content in transgenic tobacco was up to 0.3% of the total soluble protein. Using the Vero E6 cell expansion assay and the MTT method for cell proliferation, hEGF produced by transgenic tobacco significantly stimulated Vero E6 cell expansion and proliferation, the same as commercial hEGF products.

show abstract

Expression of active human epidermal growth factor (hEGF) in tobacco plants by integrative and non-integrative systems

Cited by 59 publications

References 22 publications

A rice-based edible vaccine expressing multiple T cell epitopes induces oral tolerance for inhibition of Th2-mediated IgE responses

A rice-based edible vaccine expressing multiple T cell epitopes induces oral tolerance for inhibition of Th2-mediated IgE responses

Recombinant expression of LFchimera antimicrobial peptide in a plant-based expression system and its antimicrobial activity against clinical and phytopathogenic bacteria

Expression and characteristic of synthetic human epidermal growth factor (hEGF) in transgenic tobacco plants

Contact Info

Product

Resources

About