Purification of transgenic plant-derived recombinant human acetylcholinesterase-R
Purification of transgenic plant-derived recombinant human acetylcholinesterase-R
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DOI：
10.1016/j.cbi.2005.10.097
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发表时间：
2005-12-15
影响因子：
5.1
通讯作者：
Mor, TS
中科院分区：
文献类型：
--
作者：
Geyer, BC;Muralidharan, M;Mor, TS
本拟烟（Nicotiana benthamiana）植物表达了一个密码子优化基因，该基因编码人类乙酰胆碱酯酶r （AChE）亚型。转基因植株表达该蛋白的量为可溶性蛋白总量的0 ~ 0.4%，植株产生的酶被纯化至均匀性。经裂解、普鲁卡因酰胺亲和层析和阴离子交换层析，达到400倍以上的纯化，获得电泳纯度。这种纯蛋白质在动力学上与人类乙酰胆碱酯酶的唯一商业来源难以区分，这是在哺乳动物细胞培养中产生的。因此，我们已经证明了一种生产乙酰胆碱酯酶的模型系统，它不受数量限制或与从哺乳动物细胞培养或人类血清中纯化相关的哺乳动物病原体的影响。2005爱思唯尔爱尔兰有限公司版权所有。
Nicotiana benthamiana plants were engineered to express a codon-optimized gene encoding the human acetylcholinesterase-R (AChE) isoform. The transgenic plants expressed the protein at > 0.4% of total soluble protein, and the plant-produced enzyme was purified to homogeneity. Following lysis, procainamide affinity chromatography and anion-exchange chromatography, more than 400-fold purification was achieved and electrophoretic purity was obtained. This pure protein is kinetically indistinguishable from the only commercially available source of human acetylcholinesterase, which is produced in mammalian cell culture. Thus, we have demonstrated a model system for the production of acetylcholinesterase, which is not susceptible to the quantitative limitations or mammalian pathogens associated with purification from mammalian cell culture or human serum. (c) 2005 Elsevier Ireland Ltd. All rights reserved.