Green fluorescent marker C-reactive protein in prokaryotic expression and capillary electrophoresis.docVIP

 PAGE \* MERGEFORMAT 18 Green fluorescent marker C-reactive protein in prokaryotic expression and capillary electrophoresis Author: Chen Teng-xiang, GAO Xiao-Xia, CHEN Li, Ya-Wei Liu, Liu Jinghua, Yong [Abstract] Objective: Expression and purification of recombinant protein His-EGFP-CRP by capillary electrophoresis of biological activity of the recombinant protein to evaluate. Methods: pET14b/EGFP-hCRP prokaryotic expression plasmid was transformed into E. coli BL21 (DE3) in the inducible expression of the recombinant protein purified by affinity chromatography, gradient dialysis, access to rehabilitation of His-EGFP-CRP protein; and use high-performance capillary electrophoresis (HPCE) technique to detect protein refolding to understand the isoelectric point of protein and activity. Results: The transformation test revealed that the plasmid pET14b/EGFP-hCRP in BL21 (DE3) can be induced in expression; through the inclusion bodies dissolved in denatured, affinity chromatography purification of high purity of His-EGFP-CRP protein; HPCE separation was found renaturation after the His-EGFP-CRP protein peak for more than one, and its electrophoretic fluid pH value below 6 easily detected, His-EGFP-CRP and THP-1 cell lysate after incubation increased the detection peak. Conclusion: The successfully in E. coli BL21 (DE3) was induced to express the recombinant protein His-EGFP-CRP, denatured by dissolving inclusion bodies and affinity chromatography purified to obtain the recombinant protein refolding after the His-EGFP-CRP isoelectric point close to at 6, with single or multi-polymer structure of the form, with binding activity associated with the intracellular molecules into the complex, can be applied to function and CRP in the study of the mechanism. [Keywords:] C-reactive protein; gene expression; protein refolding; electrophoresis, capillary [Abstract] Objective: To express the purified recombinant protein, His-EGFP-CRP, and to evaluate the feasibility of its