Development of Allergenicity and Toxicity Assessment Methods for Evaluating Transgenic Sugarcane Overexpressing Sucrose–Phosphate Synthase
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Materials
2.2. Animals and Housing
2.3. Allergenicity Assessment
2.3.1. Bioinformatics Analysis
2.3.2. Heat Stability Treatment
2.3.3. In Vitro Simulated Gastric Fluids (SGF) and Simulated Intestine Fluid (SIF) Assays
2.3.4. In Vivo Allergenicity Assay in Rats and Measurement of the Protein-Specific IgE Antibody
2.4. Acute Oral Toxicity Assessment
2.5. Statistical Analysis
3. Results and Discussion
3.1. Bioinformatics Assessment of the Protein SPS Expressed in Transgenic Sugarcane
3.2. Thermal Stability Assessment
3.3. In Vitro Simulated Gastric Fluids (SGF) and Simulated Intestine Fluid (SIF) Assessment
3.4. In Vivo Allergenicity Assay in Rats and Measurement of the Protein-Specific IgE Antibody
3.5. Acute Oral Toxicity Assessment
3.6. A Model for Food Safety Assessment of Transgenic Sugarcane
4. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Allergen | Species | Description | Sequence in FARRP * | Sequence Length | Matching Region of Contiguous Amino Acids |
---|---|---|---|---|---|
Chitinase | Zea mays L. | Food allergen | GI:260401081 | 279 | 62-GGGGGGGG-69 |
Endochitinase A | Zea mays L. | Food allergen | GI:116329 | 281 | 61-GGGGGGGG-68 |
Death of Animals | |||||
---|---|---|---|---|---|
Group Treatment | Sample Treatment | Dosage (gr/kg) | After 72 h | After 14 Days | Percentage (%) * |
I | Control | 0 | 0 | 0 | 0 |
II | Sugarcane Juice | 6.25 | 0 | 0 | 0 |
II | Sugarcane Juice | 12.5 | 0 | 0 | 0 |
IV | Sugarcane Juice | 25 | 0 | 0 | 0 |
Dosage of Sugarcane Juice | ||||
---|---|---|---|---|
Organs | 0 gr/kg | 6.25 gr/kg | 12.5 gr/kg | 25 gr/kg |
Heart | 0.37 ± 0.03 | 0.44 ± 0.03 | 0.41 ± 0.03 | 0.44 ± 0.04 |
Liver | 5.74 ± 0.68 | 6.25 ± 0.19 | 7.06 ± 0.07 | 6.80 ± 0.10 |
Spleen | 0.77 ± 0.17 | 0.79 ± 0.06 | 0.85 ± 0.02 | 0.84 ± 0.11 |
Kidneys | 1.31 ± 0.10 | 1.56 ± 0.03 | 1.29 ± 0.07 | 1.43 ± 0.12 |
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Neliana, I.R.; Sawitri, W.D.; Ermawati, N.; Handoyo, T.; Sugiharto, B. Development of Allergenicity and Toxicity Assessment Methods for Evaluating Transgenic Sugarcane Overexpressing Sucrose–Phosphate Synthase. Agronomy 2019, 9, 23. https://doi.org/10.3390/agronomy9010023
Neliana IR, Sawitri WD, Ermawati N, Handoyo T, Sugiharto B. Development of Allergenicity and Toxicity Assessment Methods for Evaluating Transgenic Sugarcane Overexpressing Sucrose–Phosphate Synthase. Agronomy. 2019; 9(1):23. https://doi.org/10.3390/agronomy9010023
Chicago/Turabian StyleNeliana, Intan Ria, Widhi Dyah Sawitri, Netty Ermawati, Tri Handoyo, and Bambang Sugiharto. 2019. "Development of Allergenicity and Toxicity Assessment Methods for Evaluating Transgenic Sugarcane Overexpressing Sucrose–Phosphate Synthase" Agronomy 9, no. 1: 23. https://doi.org/10.3390/agronomy9010023