AM79 EPSPS蛋白的体外模拟胃肠液消化稳定性研究
发布时间：2018-08-22 07:32
【摘要】：AM79 EPSPS基因是从草甘膦高度污染的土壤中克隆的、具有我国自主知识产权的一个新型抗草甘膦基因,在转基因玉米中过表达能显著提高转基因玉米的草甘膦抗性,具有重要应用价值。为了评价AM79 EPSPS蛋白的食用安全性,本研究参考中华人民共和国国家标准,开展了AM79 EPSPS蛋白的消化稳定性研究。将AM79 EPSPS基因构建到原核表达载体pET-30a上,在大肠杆菌中表达了分子量为48.3 kD的可溶性AM79 EPSPS蛋白。通过Ni-NTA亲和层析方法纯化了AM79 EPSPS蛋白用于体外模拟胃肠液实验。结果表明,AM79 EPSPS蛋白在模拟胃肠液中均在15 s内即被消化,SDS-PAGE和Western蛋白免疫印迹均未检测到蛋白残留,表明AM79 EPSPS蛋白在模拟胃肠液体系中极易被消化。
[Abstract]:The AM79 EPSPS gene was cloned from highly contaminated soil of glyphosate and a new glyphosate resistant gene with independent intellectual property rights in China. Overexpression in transgenic maize can significantly improve glyphosate resistance in transgenic maize. It has important application value. In order to evaluate the edible safety of AM79 EPSPS protein, the digestive stability of AM79 EPSPS protein was studied with reference to the national standard of the people's Republic of China. The AM79 EPSPS gene was constructed into prokaryotic expression vector pET-30a and the soluble AM79 EPSPS protein with molecular weight of 48.3 KD was expressed in E. coli. AM79 EPSPS protein was purified by Ni-NTA affinity chromatography to simulate gastrointestinal fluid in vitro. The results showed that the protein residues of AM79 EPSPS protein were not detected by SDS-PAGE and Western Western blotting in the simulated gastrointestinal fluid within 15 seconds, indicating that AM79 EPSPS protein was easily digested in the simulated gastrointestinal fluid system.
【作者单位】： 中国农业大学农学院;中国农业科学院作物科学研究所;
【基金】：转基因新品种培育重大专项(2016ZX08003-001)
【分类号】：S513
，
本文编号：2196413
[Abstract]:The AM79 EPSPS gene was cloned from highly contaminated soil of glyphosate and a new glyphosate resistant gene with independent intellectual property rights in China. Overexpression in transgenic maize can significantly improve glyphosate resistance in transgenic maize. It has important application value. In order to evaluate the edible safety of AM79 EPSPS protein, the digestive stability of AM79 EPSPS protein was studied with reference to the national standard of the people's Republic of China. The AM79 EPSPS gene was constructed into prokaryotic expression vector pET-30a and the soluble AM79 EPSPS protein with molecular weight of 48.3 KD was expressed in E. coli. AM79 EPSPS protein was purified by Ni-NTA affinity chromatography to simulate gastrointestinal fluid in vitro. The results showed that the protein residues of AM79 EPSPS protein were not detected by SDS-PAGE and Western Western blotting in the simulated gastrointestinal fluid within 15 seconds, indicating that AM79 EPSPS protein was easily digested in the simulated gastrointestinal fluid system.
【作者单位】： 中国农业大学农学院;中国农业科学院作物科学研究所;
【基金】：转基因新品种培育重大专项(2016ZX08003-001)
【分类号】：S513
，
本文编号：2196413
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