Journal of China Pharmaceutical University 2017;48(2):214-219
doi:10.11665/j.issn.1000-5048.20170213
Studies on preparation and properties of Staphylococcus nuclease
Liu HONG ; Junchao LANG ; Dongmei HE ; Kunfeng LIU ; Jie WU
Keywords
diabetes mellitus; Staphylococcal nuclease; His-tag; separation and purification
Country
China
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.
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