Expression of Biologically Active Rat Prolactin in Mammalian COS-1 Cells.
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- SHIMOKAWA NORIAKI
- Hormone Assay Center, Institute of Endocrinology, Gunma University
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- KATO YUKIO
- Hormone Assay Center, Institute of Endocrinology, Gunma University
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- WAKABAYASHI KATSUMI
- Hormone Assay Center, Institute of Endocrinology, Gunma University
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抄録
Rat prolactin (PRL) cDNA was constructed in mammalian expression vector, pSVL. Transient expression of rat PRL was performed in COS-1 cells by theDEAE-dextran method. The production of recombinant rat PRL started within 48h from the cells and reached the level of 1.0-1.5μg/ml/5×105 cells. Themolecular size of recombinant rat PRL was the same as that of standard ratPRL (Mw: 23, 000), suggesting successful removal of the signal peptide. Theradioimmunoassay and isoelectric focusing analysis showed that recombinant ratPRL has almost the same immunological and biochemical characteristics asthose of standard rat PRL. As biological tests, receptor-binding activity, Nb 2node lymphoma cell growth activity, and mammary gland stimulating activitywere examined. The radioreceptor assay showed that recombinant rat PRL hasbinding activity to mammary microsomal membrane similar to that of standardrat PRL. Recombinant rat PRL also stimulated the growth of Nb 2 lymphomacells as standard rat PRL. Finally it was shown that recombinant rat PRLpromotes the synthesis of the secretory materials in the lumen of mousemammary gland with the same potency as that of standard rat PRL. In conclusion, recombinant rat PRL, which was produced in mammalian cells in thepresent experiment, has immunological, biochemical and biologicalcharacteristics similar to those of standard PRL, and has full bioactivity.
収録刊行物
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- Ｅｎｄｏｃｒｉｎｏｌｏｇｉａ Ｊａｐｏｎｉｃａ
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Ｅｎｄｏｃｒｉｎｏｌｏｇｉａ Ｊａｐｏｎｉｃａ 37 (1), 141-150, 1990
一般社団法人 日本内分泌学会