Method for production of lymphotoxin (TNFB) in cell line A-C5-8

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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4351722, 43524026, 4353201, 424 851, 520351, 935 10, A61K 4505, C12P 2106, C12N 1500

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054037255

ABSTRACT:
Lymphotoxin-producing human T-cell hybridomas are incubated in a medium containing phorbol myristate acetate, concanavalin-A or a mixture thereof, the resulting cells are fractionated by sucrose density-gradient centrifugation method to isolate a messenger RNA in 12.6S to 14.6S fractions, and a gene containing a part encoding a polypeptide having a lymphotoxin-activity is prepared from the messenger RNA. The gene is represented by the nucleotide sequence of the Table 1 as herein given. Using the present gene, a new lymphotoxin having the amino acid sequence (I) or (II) as given herein can be obtained by genetic engineering technology.

REFERENCES:
Asada et al. Cell. Immunology 1983 pp. 150-160 vol. 77 "Human T-Cell Hybridomas Producing Lymphokines".
Wallach et al. FEBS vol. 178(2) Dec. 1984 "Translation of mRNA for human lymphotoxin in microinjected Xenopus oocytes".
Gray et al. Nature vol. 312 Dec. 1984 pp. 721-727 "Cloning and expression of cDNA for human lymphotoxin, a lymphokine with tumor necrosis activity".
Kobayashi et al. J. Biochem. vol. 100 1986 pp. 727-733.
Journal of Biochemistry, vol. 100, pp. 727-733 (1986).

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