Expression of recombinant anticoagulant hirudin in the differentiated cultures of the porcine mammary epithelial cell line SI-PMEC.
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.
Sun
Y L
YL
Division of Biotechnology, Animal Technology Institute, Taiwan, P.O. Box 23, Chunan 35099, Miaoli, Taiwan.
Chou
Y C
YC
Kuan
T C
TC
Tu
C F
CF
Lin
C S
CS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005343:Fibrinolytic Agents / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D006629:Hirudins / Q000096:biosynthesis* / Q000235:genetics; D007797:Laminin / Q000494:pharmacology; D008321:Mammary Glands, Animal; D008969:Molecular Sequence Data; D011388:Prolactin / Q000494:pharmacology; D011509:Proteoglycans / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013552:Swine; D014162:Transfection
D004338:Drug Combinations; D005343:Fibrinolytic Agents; D006629:Hirudins; D007797:Laminin; D011509:Proteoglycans; D011994:Recombinant Proteins; C059485:matrigel; D011388:Prolactin; D003094:Collagen