發(fā)布時(shí)間：2018-08-22 07:32

【摘要】：AM79 EPSPS基因是從草甘膦高度污染的土壤中克隆的、具有我國(guó)自主知識(shí)產(chǎn)權(quán)的一個(gè)新型抗草甘膦基因,在轉(zhuǎn)基因玉米中過(guò)表達(dá)能顯著提高轉(zhuǎn)基因玉米的草甘膦抗性,具有重要應(yīng)用價(jià)值。為了評(píng)價(jià)AM79 EPSPS蛋白的食用安全性,本研究參考中華人民共和國(guó)國(guó)家標(biāo)準(zhǔn),開(kāi)展了AM79 EPSPS蛋白的消化穩(wěn)定性研究。將AM79 EPSPS基因構(gòu)建到原核表達(dá)載體pET-30a上,在大腸桿菌中表達(dá)了分子量為48.3 kD的可溶性AM79 EPSPS蛋白。通過(guò)Ni-NTA親和層析方法純化了AM79 EPSPS蛋白用于體外模擬胃腸液實(shí)驗(yàn)。結(jié)果表明,AM79 EPSPS蛋白在模擬胃腸液中均在15 s內(nèi)即被消化,SDS-PAGE和Western蛋白免疫印跡均未檢測(cè)到蛋白殘留,表明AM79 EPSPS蛋白在模擬胃腸液體系中極易被消化。
[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.
【作者單位】： 中國(guó)農(nóng)業(yè)大學(xué)農(nóng)學(xué)院;中國(guó)農(nóng)業(yè)科學(xué)院作物科學(xué)研究所;
【基金】：轉(zhuǎn)基因新品種培育重大專項(xiàng)(2016ZX08003-001)
【分類號(hào)】：S513
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本文編號(hào)：2196413