摘要
【目的】为了获得较高活性的螨AChE重组蛋白。【方法】将叶螨ace基因全长和去疏水区两种形式分别在pColdⅡ与pET-30a载体中表达,比较它们的表达效果。【结果】AChE在两种载体中成功表达,得到了较高纯度的AChE。与载体pET-30a比,AChE蛋白在pColdⅡ表达载体中的表达量、酶活性以及对毒扁豆碱的敏感性都更高,去掉羧基端疏水区有助于提高AChE的活性。【结论】此研究建立高活性AChE的表达系统,获得了大量活性较高的AChE重组蛋白,为AChE抑制剂的筛选提供试验基础。
【Objective】The aim of this study is to obtain mite AChE protein with high activity.【Methods】The ace gene with or without 3’ hydrophobic region were expressed in expression vectors of pCold II and pET-30 a respectively, and their expression effects were compared.【Results】Various AChEs were all successfully expressed and purified from the pCold II and pET-30 a vectors.The expression levels, enzyme activities, and AChE sensitivities to eserine were higher in pCold II than that in pET-30 a vector.The deletion of the 3’ hydrophobic region of ace gene improved the activity of AChE protein.【Conclusion】In this study, the in vitro expression system of high active AChE was constructed, and highly active recombinant AChE protein in quantity has obtained.This study laid the foundation for screening acetylcholinesterase inhibitors.
作者
韦显星
李博
张翼鹏
丁超
杨金
彭博
李梦怡
卜春亚
WEI Xianxing;LI Bo;ZHANG Yipeng;DING Chao;YANG Jin;PENG Bo;LI Mengyi;BU Chunya(College of Biological and Resource Environment/Key Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs,Beijing University of Agriculture,Beijing 102206,China)
出处
《北京农学院学报》
2020年第4期9-14,共6页
Journal of Beijing University of Agriculture
基金
国家自然科学基金项目(31670648)
北京市自然科学基金项目(6162004)
教育部科学技术重点研究项目(212001)。
