Summary
We have constructed the recombinant plasmid for the extracellular production of human immunoglobulin G Fc region (hIgG-Fc) in Escherichia coli. The excretion vector pEXFC10 contained the weakly activated kil gene of plasmid pMB9 and the DNA fragment encoding a fused protein, in which the codons for the alkalophilic Bacillus sp. No. 170 penicillinase signal peptide and the hIgG-Fc were fused through the one additional amino acid Ser, which was identical with the N-terminus of alkalophilic Bacillus mature penicillinase. By cultivating E. coli carrying pEXFC10, about 40% of hIgG-Fc was excreted into the culture medium. The N-terminal amino acid sequence of the extracellular hIgG-Fc indicated that processing occurred correctly between Ala and Ser. From the sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE) in the nonreducing condition, it was suggested that most of the extracellular hIgG-Fc proteins took the dimeric form via disulfide bonds.
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Kitai, K., Kudo, T., Nakamura, S. et al. Extracellular production of human immunoglobulin G Fc region (hIgG-Fc) by Escherichia coli . Appl Microbiol Biotechnol 28, 52–56 (1988). https://doi.org/10.1007/BF00250497
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DOI: https://doi.org/10.1007/BF00250497