Hyperproduction of human interferon γ by rat cells maintained in low-serum medium using the fibronectin gene promoter

Takuma Nakajima, Shiho Tsunoda, Susumu Nakada, Shigekazu Nagata, Kinichiro Oda

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

An expression vector, pF1900M, which expresses a cloned gene at a high level in quiescent mammalian cells was constructed using the rat fibronectin (FN) promoter. Human interferon γ (HuIFN- γ) cDNA inserted downstream of the FN promoter in pF1900M was introduced into rat 3Y1 cells and several IFN-producing cell lines were established. These cells secreted a low level of IFN when they were growing but secreted at a high level after they had reached confluence. The level was further increased when the confluent cells were maintained in low-serum medium and a cell line, I7, produced 4×105 IU/ml of IFN, comparable to that produced by genetically engineered Escherichia coli in 2 days. The IFN-producing ability of 17 cells could be maintained by successive replacements of low-serum medium for at least 2 weeks. HuIFN- γ secreted into the medium had a molecular weight range of 22,000 to 25,000, similar to that of IFN- γ produced by human lymphocytes. The N-linked glycosylation of HuIFN- γ seemed to occur properly, since treatment of the IFN with N-glycanase resulted in a reduction of molecular weight to 17,000, which corresponds to that calculated from the deduced amino acid sequence of HuIFN-γ.

Original languageEnglish (US)
Pages (from-to)590-597
Number of pages8
JournalJournal of Biochemistry
Volume112
Issue number5
DOIs
StatePublished - Jan 1 1992

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Serum-Free Culture Media
Fibronectins
Interferons
Rats
Genes
Cells
Molecular weight
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Glycosylation
Molecular Weight
Lymphocytes
Cell Line
Escherichia coli
Complementary DNA
Amino Acid Sequence
Amino Acids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Hyperproduction of human interferon γ by rat cells maintained in low-serum medium using the fibronectin gene promoter. / Nakajima, Takuma; Tsunoda, Shiho; Nakada, Susumu; Nagata, Shigekazu; Oda, Kinichiro.

In: Journal of Biochemistry, Vol. 112, No. 5, 01.01.1992, p. 590-597.

Research output: Contribution to journalArticle

Nakajima, Takuma ; Tsunoda, Shiho ; Nakada, Susumu ; Nagata, Shigekazu ; Oda, Kinichiro. / Hyperproduction of human interferon γ by rat cells maintained in low-serum medium using the fibronectin gene promoter. In: Journal of Biochemistry. 1992 ; Vol. 112, No. 5. pp. 590-597.
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