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目的评价转G10evo、Cry1Ab/Cry2Ab基因抗虫耐草甘膦玉米(GAB-3)中的外源基因表达蛋白1Ab、2Ab和G10(EPSPS)在模拟胃肠液中的消化稳定性。方法根据国家标准建立体外模拟胃肠环境消化体系,对照或样品均以5 g/L进行模拟胃液消化,以2 g/L进行模拟肠液消化。于反应0 s、15 s、2 min、30 min和60 min时取出200μl模拟消化液,加入上样缓冲液并沸水浴5 min终止反应。然后将消化后的对照及样品进行十二烷基磺酸钠-聚丙烯酰胺凝胶(SDS-PAGE)电泳、染色并使用凝胶成像系统观察结果。结果作为稳定对照的大豆胰蛋白酶抑制剂(soybean trypsin inhibitor,STI)在模拟胃肠液中60 min未被消化,作为不稳定对照的酪蛋白(α-casein)在模拟胃肠液中15 s内全部消化。外源基因表达蛋白1Ab、2Ab和G10(EPSPS)在模拟胃液中2 min内完全消化,在模拟肠液中30 min内完全消化。结论转基因玉米(GAB-3)的外源基因表达蛋白1Ab、2Ab和G10(EPSPS)在模拟胃肠液中不具有消化稳定性,容易被降解。
Objective To evaluate the digestibility of exogenous gene expression proteins 1Ab, 2Ab and G10 (EPSPS) in transgenic G10evo, Cry1Ab / Cry2Ab resistant and glyphosate tolerant maize (GAB-3) in simulated gastrointestinal fluid. Methods According to the national standard, a gastrointestinal digestive system was established in vitro. The control or samples were all digested with simulated gastric juice at 5 g / L and simulated intestinal fluid at 2 g / L. At 0 s, 15 s, 2 min, 30 min and 60 min, 200 μl of simulated digestion solution was removed, and the reaction was stopped by adding a loading buffer and boiling water bath for 5 min. The digested controls and samples were then subjected to sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) electrophoresis, stained and observed using a gel imaging system. Results As a stable control, soybean trypsin inhibitor (STI) was not digested in simulated gastrointestinal fluid for 60 min, while casein (α-casein) as an unstable control in simulated gastrointestinal fluid within 15 s All digestion. Exogenous gene expression proteins 1Ab, 2Ab and G10 (EPSPS) were completely digested within 2 min in simulated gastric juice and completely digested in simulated intestinal fluid within 30 min. Conclusion The exogenous gene expression proteins 1Ab, 2Ab and G10 (EPSPS) of transgenic maize (GAB-3) are not digested and stable in the simulated gastrointestinal fluid and are easily degraded.