玉米赤霉烯酮降解酶基因zhd101表达载体构建及在大肠杆菌中表达

发布时间：2018-01-11 01:27

本文关键词：玉米赤霉烯酮降解酶基因zhd101表达载体构建及在大肠杆菌中表达　出处：《畜牧与兽医》2016年01期 　论文类型：期刊论文

更多相关文章： 玉米赤霉烯酮 zhd基因 大肠杆菌 表达

【摘要】：人工合成玉米赤霉烯酮降解酶基因zhd101,以质粒p ET32a(+)为表达载体,构建重组质粒,转化大肠杆菌BL21(DE3),以IPTG诱导转化子,通过SDS-PAGE鉴定ZHD101蛋白表达水平,并纯化ZHD101蛋白。结果表明,重组质粒p ET32a(+)-zhde101经HindⅢ、SacⅠ酶切后,1%琼脂糖凝胶电泳证实zhd101正确插入p ET32a(+)载体。重组菌经IPTG诱导后,表达的ZHD101蛋白分子量为47.5 ku,蛋白表达量约占细菌总蛋白量的35%,纯化后的ZHD101蛋白样品经SDS-PAGE和光密度扫描分析表明其纯度为95%左右。试验结果为进一步研究ZHD101蛋白功能奠定了基础。
[Abstract]:The artificial synthesis of zearalenone degradation enzyme gene zhd101, plasmid P ET32a (+) as the expression vector, the recombinant plasmid was constructed and transformed into Escherichia coli BL21 (DE3), the transformants were induced by IPTG, the expression level of ZHD101 protein identified by SDS-PAGE, and purified ZHD101 protein. The results show that the recombinant plasmid P (+ ET32a -zhde101) by Hind III, Sac I digestion, 1% agarose gel electrophoresis confirmed that zhd101 ET32a is correctly inserted into P (+) vector. The recombinant bacteria induced by IPTG, ZHD101 protein expression was 47.5 Ku, the expression of about 35% of the amount of total bacterial protein, the purified ZHD101 protein samples by SDS-PAGE and optical density scanning analysis showed that the purity was about 95%. The results laid the foundation for further study of ZHD101 protein function.

【作者单位】：沈阳农业大学畜牧兽医学院;四川理工学院化学与制药工程学院;沈阳农业大学植物保护学院;
【基金】：国家自然科学基金(31201961;31302152) 中国博士后科学基金资助(2014M551125) 辽宁省教育厅科学研究一般项目(L2014561)
【分类号】：S859.8;S816
【正文快照】： 镰刀菌产生的玉米赤霉烯酮(zearalenone,ZEN)是世界上污染最为广泛的一种毒素[1],具有很强的雌激素样作用及生殖发育毒性[2]、免疫毒性[3],如何对该毒素脱毒的研究受到世界各国的普遍重视。传统的ZEN去毒方法有物理和化学方法,包括氨化法、碱法、高温法、紫外线照射法、超滤-

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