Extracellular Production of Human Tumor Necrosis Factor-α by Escherichia coli Using a Chemically-synthesized Gene(Biological Chemistry)

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A DNA fragment of approximately 490 base pairs encoding human TNF was chemically synthesized and expressed within Escherichia coli cells. Furthermore, extracellular production of human TNF and several N-terminal deletion mutants of TNF was attempted using the excretion vector pEAPS. The TNF mutant with two N-terminal amino acids deleted (NΔ2-TNF) was efficiently excreted into the culture medium by E. coli carrying the plasmid pEXTNF3. In this clone, the signal peptide was correctly processed during the excretion. The E. coli-excreted NΔ2-TNF had higher antitumor activity than wild-type TNF or NΔ2-TNF produced intracellularly by E. coli.
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