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Development of Allergenicity and Toxicity Assessment Methods for Evaluating Transgenic Sugarcane Overexpressing Sucrose–Phosphate Synthase

机译:评价过表达蔗糖-磷酸合酶的转基因甘蔗的致敏性和毒性评估方法的发展

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Sugarcane is considered as an industrial crop that produces sugar. The number of transgenic sugarcane on the market is currently increasing. Therefore, investigation of the potential allergens and toxics in transgenic sugarcane is necessary, since there is less information regarding food safety for human consumption. Bioinformatics and experimental analysis were used for the validation of the allergenic potential of transgenic sugarcane overexpressing sucrose–phosphate synthase (SPS). Bioinformatics analysis showed that SPS has no homology with any known allergenic proteins. However, eight-residues identical contiguous sequence was detected, and further specific assessment is required to confirm the potential of allergenicity. The results of protein stability evaluation showed that SPS gradually decreased at 28 °C and rapidly inactivated at 60 °C and 90 °C by heat treatment. In addition, total protein was degraded by simulated gastric fluids (SGF), and simulated intestine fluid (SIF) assays for one-minute incubation. The level of specific IgE in the transgenic sugarcane and controls also showed no potential risk of allergy. An acute oral toxicity assay was performed by oral gavage of transgenic sugarcane juice in mice. The LD 50 for transgenic sugarcane juice was 25 gr/kg body weight. We propose a development method for allergenicity and toxicity assessment in transgenic sugarcane.
机译:甘蔗被认为是生产糖的工业作物。目前市场上转基因甘蔗的数量正在增加。因此,有必要对转基因甘蔗中的潜在过敏原和毒性进行研究,因为关于人类食用食品安全的信息较少。生物信息学和实验分析被用于验证过表达蔗糖-磷酸合酶(SPS)的转基因甘蔗的致敏潜力。生物信息学分析表明,SPS与任何已知的致敏蛋白均无同源性。但是,检测到八个残基相同的连续序列,需要进一步的具体评估以确认潜在的致敏性。蛋白质稳定性评估的结果显示,通过热处理,SPS在28°C时逐渐降低,并在60°C和90°C时迅速失活。此外,总蛋白会通过模拟胃液(SGF)和模拟肠液(SIF)分析降解一分钟。转基因甘蔗和对照中特异性IgE的水平也没有显示过敏的潜在风险。通过口服灌胃转基因甘蔗汁对小鼠进行急性口服毒性试验。转基因甘蔗汁的LD 50> 25 gr / kg体重。我们提出了一种开发方法,用于转基因甘蔗的致敏性和毒性评估。

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