Studies on preparation and properties of Staphylococcus nuclease

HONG Liu; LANG Junchao; HE Dongmei; LIU Kunfeng; WU Jie

doi:10.11665/j.issn.1000-5048.20170213

Journal of China Pharmaceutical University > 2017 > 48(2): 214-219. > DOI: 10.11665/j.issn.1000-5048.20170213

HONG Liu, LANG Junchao, HE Dongmei, LIU Kunfeng, WU Jie. Studies on preparation and properties of Staphylococcus nuclease[J]. Journal of China Pharmaceutical University, 2017, 48(2): 214-219. DOI: 10.11665/j.issn.1000-5048.20170213

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Studies on preparation and properties of Staphylococcus nuclease

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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 His-SNase 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.
- diabetes mellitus,
- Staphylococcal nuclease,
- His-tag,
- separation and purification

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