Expression of recombinant anticoagulant hirudin in the differentiated cultures of the porcine mammary epithelial cell line SI-PMEC

Cell Biology International
Y L SunC S Lin

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.

Citations

Nov 14, 2013·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Tang-Ching KuanChih-Sheng Lin
Aug 11, 2018·Biotechnology & Genetic Engineering Reviews·Jianguo Zhang, Nana Lan
Nov 15, 2020·Scientific Reports·Doreen A WüstenhagenStefan Kubick

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