军事医学2016，Vol.40Issue(4)：319-321,3.DOI:10.7644/j.issn.1674-9960.2016.04.013
A型肉毒毒素轻链多步纯化及其金属蛋白酶活性研究
Multi-step purifications of botulinum neurotoxin A light chain and identification of its metalloproteases activity
摘要
Abstract
Objective To obtain highly purified botulinum neurotoxin A light chain(BoNT-ALC) protein in E.coli by genetic engineering and multi-step purifications, and identify its metalloproteases activity.Methods The full-length of BoNT-ALC was cloned from BoNT A by PCR and inserted into plasmid pET-22b.Then pET-22b-ALC was transformed into E.coli BL21( DE3) strains and induced by IPTG.The protein was purified by Ni-NTA sepharose,anion exchange column and gel filtration.The enzymatic activity of the protein was identified by SNAP-25.Results and Conclusion A highly purified and homogeneous protein is obtained, which shows good enzymatic activity.关键词
A型肉毒毒素/蛋白纯化/金属蛋白酶类Key words
botulinum neurotoxin A/protein purification/metalloproteases分类
医药卫生引用本文复制引用
卢晓雪,赵缓,黄彦杰,李涛,王慧..A型肉毒毒素轻链多步纯化及其金属蛋白酶活性研究[J].军事医学,2016,40(4):319-321,3.基金项目
国家自然科学基金资助项目 （）
