Isolation and Characterization of Human Prorenin Secreted from Murine Cells Transformed with a Bovine Papilloma Virus–Preprorenin Expression Vector

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Abstract

We report the construction of a plasmid–based expression vector that carries the murine metallothionein gene promoter, the human preprorenin gene, the Tn5 phosphotransferase gene, and a complete bovine papilloma virus genome. Murine cells transformed with this vector constitutively secrete high levels of human prorenin as determined by immunoprecipitation of culture media with anti–human renin antibody and activity assays. An immunoaffinity system for the isolation of human prorenin from serum–free media, or media containing serum, was developed. Purified human prorenin is stable for months and is fully activated to enzymatically mature renin by limited tryptic digestion. This is the first example of a recombinant system leading to the isolation of research quantities of highly pure and fully activatable human prorenin.

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Evans, D., Weighous, T., Cornette, J. et al. Isolation and Characterization of Human Prorenin Secreted from Murine Cells Transformed with a Bovine Papilloma Virus–Preprorenin Expression Vector. Nat Biotechnol 5, 705–709 (1987) doi:10.1038/nbt0787-705

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