Expression of recombinant anticoagulant hirudin in the differentiated cultures of the porcine mammary epithelial cell line SI-PMEC |
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Authors: | Sun Y L Chou Y C Kuan T C Tu C F Lin C S |
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Affiliation: | Division of Biotechnology, Animal Technology Institute, Taiwan, P.O. Box 23, Chunan 35099, Miaoli, Taiwan. |
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Abstract: | 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 beta-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 0.5-0.6microg 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. |
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Keywords: | Recombinant protein Mammary gland Epithelial cell SI‐PMEC Hirudin |
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