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Amplification and High-Level Expression of a cDNA for Human Granulocyte-Macrophage Colony-Stimulating Factor in Human Lymphoblastoid Namalwa Cells

Bio/Technology volume 8, pages 550–553 (1990)Cite this article

Abstract

We have achieved high-level expression of human granulocyte-macrophage colony-stimulating factor (GM-CSF) in human lymphoblastoid Namalwa cells by introducing and subsequently amplifying an expression vector encoding human GM-CSF and mouse dihydrofolate reductase (DHFR), Transformants expressing elevated levels of both GM-CSF and DHFR were selected by a step-wise increase in the concentration of methotrexate (MTX). Several cell lines resistant in 800nM MTX have been established that express GM-CSF at a rate of 10–20 μg/106 cells/day. The amplified genes are integrated and stably maintained in the chromosomes of these cell lines. Analysis of the GM-CSF produced in the amplified Namalwa cells revealed that the molecular-size distribution due to varied degrees of glycosylation was similar to that observed in the naturally-occurring molecules.

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Author notes

  1. Hideki Yanagi: Corresponding author.

Authors and Affiliations

  1. Biotechnology Laboratory, Takarazuka Research Center, Sumitomo Chemical Co., Ltd., 4-2-1 Takatsukasa, Takarazuka, Hyogo, 665, Japan

    Minoru Okamoto, Chikao Nakayama, Michio Nakai & Hideki Yanagi

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  1. Minoru Okamoto
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  4. Hideki Yanagi
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Okamoto, M., Nakayama, C., Nakai, M. et al. Amplification and High-Level Expression of a cDNA for Human Granulocyte-Macrophage Colony-Stimulating Factor in Human Lymphoblastoid Namalwa Cells. Nat Biotechnol 8, 550–553 (1990). https://doi.org/10.1038/nbt0690-550

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