Extracellular production of human tumor necrosis factor-alpha by Escherichia coli using a chemically-synthesized gene.
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 pEAP8. The TNF mutant with two N-terminal amino acids deleted (N delta 2-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 N delta 2-TNF had higher antitumor activity than wild-type TNF or N delta 2-TNF produced intracellularly by E. coli.
Biotechnology Research Laboratories, Teijin Limited, Tokyo, Japan.
S
S
Nakamura
T
T
Masegi
K
K
Kitai
Y
Y
Ichikawa
T
T
Kudo
R
R
Aono
K
K
Horikoshi
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005813:Genes, Synthetic; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012513:Sarcoma, Experimental / Q000188:drug therapy; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000737:chemistry / Q000627:therapeutic use
D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha