Studies on preparation and properties of Staphylococcus nuclease
10.11665/j.issn.1000-5048.20170213
- VernacularTitle:葡萄球菌核酸酶的制备及性质研究
- Author:
Liu HONG
;
Junchao LANG
;
Dongmei HE
;
Kunfeng LIU
;
Jie WU
- Keywords:
diabetes mellitus;
Staphylococcal nuclease;
His-tag;
separation and purification
- From:
Journal of China Pharmaceutical University
2017;48(2):214-219
- CountryChina
- Language:Chinese
-
Abstract:
In order to study the relationship between Staphylococcal nuclease(SNase) and diabetes mellitus,genetic engineering bacteria E.coli BL21/pET28a-His-SNase was constructed,the expression of soluble extracellular protein SNase was induced and the a preliminary research was made on it.An expression vector pET28a-His-SNase plasmid containing the His-SNase gene was constructed and transformed into E.coli BL21 (DE3) competent cells.The protein was induced by lactose and purified by ultrasound destruction and Ni-affinity chromatography,respectively.It was then analyzed by SDS-PAGE and Western blot.The enzymatic properties for SNase has been preliminary studied as well.Results indicated that the purity of the correctly expressed fusion protein HisSNase was over 85％.SNase showed good activity within a wide range of pH and good heat resistance.This experiment might be a foundation work for the further study on the relationship between SNase and with diabetes.
