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Volume 271, Number 28, Issue of July 12, 1996 pp. 16962-16966
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Recombinant Protein Synthesis in Chinese Hamster Ovary Cells Using a Vaccinia Virus/Bacteriophage T7 Hybrid Expression System

(Received for publication, February 13, 1996, and in revised form, April 24, 1996)

From the Laboratory of Viral Diseases, NIAID, National Institutes of Health, Bethesda, Maryland 20892

The vaccinia virus/bacteriophage T7 expression system was adapted to Chinese hamster ovary (CHO) cells. Vaccinia virus undergoes abortive infection in CHO cells, which is characterized by a sharp reduction in protein synthesis at the stage of viral intermediate gene expression. We determined that expression of a T7 promoter-regulated chloramphenicol acetyltransferase gene was at least 20 times more efficient in permissive BS-C-1 than in CHO cells. The encephalomyocarditis virus 5-untranslated region, which confers cap-independent translatability to mRNA, stimulated recombinant protein synthesis by 10-fold in both cell lines, maintaining the advantage of the BS-C-1 cells over CHO cells. Since the cowpox virus hr gene overcomes vaccinia virus host range restriction in CHO cells, we constructed a recombinant virus that carries an intact hr gene in addition to the T7 RNA polymerase gene. With this virus, synthesis of T7 RNA polymerase was enhanced and production of a recombinant protein occurred in CHO cells at the level observed in permissive cell lines. Extension of the vaccinia virus/bacteriophage T7 expression system to CHO cells should be of wide interest, as these cells have advantages for preparation of recombinant proteins in research and biotechnology.

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