The Expression and Immunoreactive Analysis of Recombinant Human C-reactive Protein in Different Prokaryotic Expression Vectors and Strains

doi:10.13560/j.cnki.biotech.bull.1985.2016.03.026

Abstract

Abstract: The prokaryotic expression vector of human C-reactive protein(CRP)was constructed and transferred into Escherichia coli BL21(DE3)and Rosetta gami2(DE3)pLysS, respectively, and then these recombinant strains were induced for the expression of CRP. Mainly gene engineering, affinity chromatography and dialysis refolding methods were employed. The expression vectors and recombinant strains of human CRP were constructed successfully, and the varied CRPs with different protein tags were acquired by IPTG. In conclusion, expressions of the same vector in different strains differed；it expressed in the E. coli BL21(DE3)and Rosetta gami2(DE3)pLysS respectively, and the inclusion bodies of recombinant CRP were obtained. The results of detecting the refolded recombinant CRP by Western blotting and ELISA revealed that the immunoreactivity of recombinant CRP in the prokaryotic expression vector was low, and CRP may have the immunoreactivity while it is in the pentamer form.

Key words: C-reactive protein, prokaryotic expression, inclusion bodies, refolding.

LI Jiang-feng, JIAO Li-yuan, ZHAO Xiao-ni, WANG Ji-hua, CAI Lei. The Expression and Immunoreactive Analysis of Recombinant Human C-reactive Protein in Different Prokaryotic Expression Vectors and Strains[J]. Biotechnology Bulletin, 2016, 32(3): 166-170.

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