摘要
目的评价转G2-EPSPS和GAT基因耐草甘膦大豆中的外源基因重组蛋白G2-EPSPS和GAT在模拟胃肠液中的消化稳定性。方法建立体外模拟胃肠环境的消化体系,样品或对照蛋白以5 g/L的浓度进行模拟胃液消化,以2 g/L的浓度进行模拟肠液消化。分别于反应0 s、15 s、2 min、30 min和60 min时取200μl模拟消化液,加入上样缓冲液并沸水浴5 min终止反应,然后取样进行十二烷基硫酸钠-聚丙烯酰胺凝胶(SDS-PAGE)电泳,并进行蛋白印迹(Western blot)分析。结果G2-EPSPS和GAT重组蛋白在模拟胃、肠液中均在0~15 s内完全消化,SDS-PAGE电泳与Western blot未检测到蛋白残留。结论耐草甘膦转基因大豆的外源基因重组蛋白G2-EPSPS和GAT在模拟胃肠液体系中极易被消化。
Objective To understand the digestive stability of recombinant G2-EPSPS and GAT proteins of glyphosate resistant transgenic soybean in simulated gastric and intestinal fluid.Methods An in vitro model of simulated gastrointestinal digestion system was established according to the national standard.The concentrations of proteins in simulated gastric fluid and intestinal fluid were 5 g/L and 2 g/L respectively.200μl of simulated digestive solution was taken out at the reaction time of0 s,15 s,2 min,30 min and 60 min respectively,and were added with loading buffer,the reaction was stopped by boiling water bath for 5 min,the proteins that had been digested were measured by SDS-PAGE and Western blot.Results Recombinant G2-EPSPS and GAT proteins were digested completely within 0-15 s in simulated gastric and intestinal fluid,and no protein residues were detected by SDS-PAGE and Western blot analysis.Conclusion Recombinant G2-EPSPS and GAT proteins of glyphosate resistant transgenic soybean are very easily digested in simulated gastrointestinal digestion system.
作者
秦文
王丽媛
杨倬
王晶波
卓勤
李岩
QIN Wen;WANG Li-yuan;YANG Zhuo;WANG Jing-bo;ZHUO Qin;LI Yan(Key Laboratory of Trace Element Nutrition of National Health Commission,National Institute for Nutrition and Health,Chinese Center for Disease Control and Prevention,Beijing 100050,China;不详)
出处
《环境与健康杂志》
CAS
北大核心
2020年第7期610-614,共5页
Journal of Environment and Health
基金
国家科技重大专项(2019ZX08013002)
