Expression and activity identification of recombinant human stanniocalcin 1
10.7644/j.issn.1674-9960.2014.01.011
- VernacularTitle:人斯钙素1在大肠杆菌中的重组表达与活性鉴定
- Author:
Peipei LIU
;
Chuangchuang DONG
;
Guangzhi LIU
;
Junbo SUN
;
Juxin ZHANG
- Publication Type:Journal Article
- Keywords:
stanniocalcin 1;
prokaryotic expression;
renaturation;
phosphate reabsorption
- From:
Military Medical Sciences
2014;(1):39-43
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain recombinant human stanniocalcin 1 ( STC1 ) with biological activity in Escheri.coli cells expression.Methods The gene was cloned into pET32b( +) vector by fused with thioredoxin and His tag .E.coli BL21(DE3) competent cells were transfomed by the recombinant vector .After renaturation, the fusion protein was digested with thrombin and intact STC1 protein was purified from the digested protein using Ni ion affinity chromatography .Recombi-nant humanSTC1 protein was confirmed by Western blot analysis using goat anti-STC1 antibody.The biological activity of STC1 in rat was assayed using standard method for assessment of renal function .Results The recombinant human STC 1 fu-sion protein is successfully expressed in Escherichia coli, the fusion protein was purified by affinity chromatography from the inclusion body and renaturated .Intact hSTC1 protein was released by thrombin digestion and purified by Ni ion affinity col-umn.The intact STC1 proteins was confirmed by Western blot analysis .Rat bioassay revealed that STC1 boosted phosphate reabsorption.Conclusion Recombinant STC1 protein was successfully expressed and has native biological activities .This protein could be used as an antigen for the preparation of monoclonal antibody against humanSTC 1.
