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Expression of recombinant anticoagulant hirudin in the differentiated cultures of the porcine mammary epithelial cell line SI‐PMEC
Author(s) -
Sun Y.L.,
Chou Y.C.,
Kuan T.C.,
Tu C.F.,
Lin C.S.
Publication year - 2008
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2008.02.004
Subject(s) - recombinant dna , matrigel , transfection , cell culture , hirudin , microbiology and biotechnology , biology , chemistry , cell , biochemistry , gene , immunology , thrombin , genetics , platelet
To express recombinant proteins in the spontaneously immortalized porcine mammary epithelial cell line (SI‐PMEC) currently established in our laboratory, a chemically synthesized DNA fragment encoding the anticoagulant hirudin was used to construct a mammalian expression vector under the control of the goat β‐casein regulatory sequence. The vector, named pGB562/Hi, was transfected into the SI‐PMEC cells to yield pGB562/Hi/SI‐PMEC. The pGB562/Hi/SI‐PMEC cells expressed recombinant hirudin only when they were differentiated into functional structures by growth on a Matrigel‐coated petri dish supplemented with the lactogenic hormone prolactin. The differentiated pGB562/Hi/SI‐PMEC cells produced about 0.5–0.6 μg of recombinant hirudin/mg of total cellular protein. These results suggest that the established SI‐PMEC cells have pharmaceutical potential to inducibly express bioactive heterogeneous proteins.