南京大学学报（自然科学版）2000，Vol.36Issue(4)：515-517,3.
东亚钳蝎抗神经兴奋肽在大肠杆菌中的表达
EXPRESSION OF ANTI-NEUROEXCITATION PEPTIDE (ANEP) OF SCORPION BUTHUS MARTENSII KARSCH IN ESCHERICHIA COLI
张景海 1华子春 2徐昭 1郑伟娟 1朱德煦1
作者信息
- 1. 南京大学生命科学院医药生物技术国家重点实验室,江苏,南京,210093
- 2. 沈阳药科大学生物化学教研室,辽宁,沈阳,110015
- 折叠
摘要
Abstract
According to the cDNA sequence of anti-neurcexcitation peptide of scorpion Buthus martensii Karsch, the putative mature anti-neuroexcitation peptide (ANEP) encoding DNA fragment was obtained by PCR method, and then was cloned into expression plasmid pET28a, fused with His tag at its 3' etd. When expressed in E. coli BL21 (DE3), the expression of recombinant ANEP was 15% of total cellular proteins meanwhile most recombinant ANEP products existed in the form of insoluble inclusion bodies. Co-expression of molecular chaperones or protein disulfide isomerase could not improve its solubility. The recombinant ANEP in the cell lysate was purified to homogeneity by metal chelating affinity chromatography and superdex 30 chromatography. In bioassay with convulsive mice model induced by thiosemicarbazide, recombinant ANEP could apparently delay the convulsion seizure of modal animals by 18% and showed anti-neuroexcitatory activity.关键词
东亚钳蝎/抗神经兴奋/表达Key words
Buthus martensii Karsch/anti-neuroexcitation/expression分类
生物科学引用本文复制引用
张景海,华子春,徐昭,郑伟娟,朱德煦..东亚钳蝎抗神经兴奋肽在大肠杆菌中的表达[J].南京大学学报（自然科学版）,2000,36(4):515-517,3.
