Molecular cloning of recombinant fibronectin EDA and EDB fusion protein

Abstract

Abstract: Objective Construct a recombinant plasmid pET28a-EDA-EDB,prepare the fusion EDA-EDB protein.Methods For the production of recombinant fibronectin EDA-EDB in Escherichia coli,the EDA and EDB segments were separated from pGEM2-EDA/EDB and recomposed with two additional amino acids,then cloned into the expression vector pET28a.pET system to express EDA-EDB fusion protein and6 × His/Ni-NTA system to purify it in a single step were used.Western blotting confirmed the purified protein.Results The EDA and EDB segments were ligated and inserted into pET28a vector.EDA-EDB fusion protein was highly expressed in Escherichia coli BL21(DE3).Afterwards,it was purified by Ni-NTA resin and verified by western blotting.Conclusion EDA-EDB fusion protein can be expressed in pET system and purified by6 × His/Ni-NTA system.

Key words: fibronectin, EDA\EDB domain, recombinant proteins

XUE AI-MIN;WANG HUA;YE RONG;ET AL.(DEPARTMENT OF FORENSIC MEDICINE;SHANGHAI MEDICAL SCHOOL;FUDAN UNIVERSITY;SHANGHAI000). Molecular cloning of recombinant fibronectin EDA and EDB fusion protein[J]. , 2002, 0(3): 140-143.

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Molecular cloning of recombinant fibronectin EDA and EDB fusion protein

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