Expression, purification and NETs degradation activity of human DNase I
10.11665/j.issn.1000-5048.20190113
- VernacularTitle:人DNase I的表达、纯化及降解NETs活性研究
- Author:
Yixuan LIANG
1
;
Jie WU
Author Information
1. 中国药科大学生命科学与技术学院酶工程实验室
- Publication Type:Journal Article
- Keywords:
deoxyribonuclease I;
neutrophil extracellular traps;
separation and purification;
His-tag
- From:
Journal of China Pharmaceutical University
2019;50(1):93-99
- CountryChina
- Language:Chinese
-
Abstract:
In order to study the effect of human deoxyribonuclease I(DNase I)on neutrophil extracellular traps(NETs)degradation, genetic engineering bacteria E. coli Rosetta(DE3)/pET32a-His-DNase I was constructed. The fusion protein His-DNase I was induced by lactose, and purified by Ni-affinity chromatography. Mouse neutrophils were extracted and stimulated with phorbol-12-myristate-13-acetate(PMA)to form NETs. The degradation activity of the fusion protein on NETs was detected by SytoxGreen and fluorescence microscopy. The results showed that the purified His-DNase I had high nuclease activity. This study provided the research foundation for further exploration of the clinical application of DNase I.
