Multi-step purifications of botulinum neurotoxin A light chain and identification of its metalloproteases activity
10.7644/j.issn.1674-9960.2016.04.013
- VernacularTitle:A型肉毒毒素轻链多步纯化及其金属蛋白酶活性研究
- Author:
Xiaoxue LU
;
Huan ZHAO
;
Yanjie HUANG
;
Tao LI
;
Hui WANG
- Publication Type:Journal Article
- Keywords:
botulinum neurotoxin A;
protein purification;
metalloproteases
- From:
Military Medical Sciences
2016;40(4):319-321
- CountryChina
- Language:Chinese
-
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.
