Molecular cloning of recombinant fibronectin EDA and EDB fusion protein / 法医学杂志

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 and 6 x 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 by 6 x His/Ni-NTA system.