Transformation of recombinant plasmid pJ404-EGFRvIII-bfp into an Escherichia coli NiCo21(DE3) expression host and its characterization

Epidermal growth factor receptor variant III (EGFRvIII) is a mutant of epidermal growth factor receptor (EGFR) that lacks 267 amino acids (exons 2-7) within its extracellular domain that results in the formation of a new epitope as a tumor specific target. A synthetic gene of EGFRvIII has been constructed by previous researchers to encode a fusion protein as a marker in targeted cancer therapies. This research was conducted to transform the recombinant plasmid pJ404-EGFRvIII into Escherichia coli NiCo21(DE3) host cells and characterize the E. coli NiCo21(DE3) transformants. Recombinant plasmid pJ404-EGFRvIII was isolated with an alkali lysis method and transformed into E. coli NiCo21(DE3) by heat-shock method. The transformants were grown on LB medium containing100 μg/ml ampicillin and characterized by colony PCR method. The results showed that the pJ404-EGFRvIII recombinant plasmid was transformed successfully into E. coli NiCo21(DE3). With the result that, EGFRvIII gene might be express by E. coli NiCo21(DE3) for further analysis of protein expression and purification in tumor terapy.