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...
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| Published in | Cell biology international Vol. 32; no. 7; pp. 739 - 747 |
|---|---|
| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Oxford, UK
Elsevier Ltd
01.07.2008
Blackwell Publishing Ltd |
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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 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. |
|---|---|
| AbstractList | 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. 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. 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. 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 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. |
| Author | Kuan, T.C. Lin, C.S. Tu, C.F. Chou, Y.C. Sun, Y.L. |
| Author_xml | – sequence: 1 givenname: Y.L. surname: Sun fullname: Sun, Y.L. organization: Division of Biotechnology, Animal Technology Institute, Taiwan, P.O. Box 23, Chunan 35099, Miaoli, Taiwan – sequence: 2 givenname: Y.C. surname: Chou fullname: Chou, Y.C. organization: Division of Biotechnology, Animal Technology Institute, Taiwan, P.O. Box 23, Chunan 35099, Miaoli, Taiwan – sequence: 3 givenname: T.C. surname: Kuan fullname: Kuan, T.C. organization: Department of Biological Science and Technology, National Chiao Tung University, No. 75 Po-Ai Street, Hsinchu 30068, Taiwan – sequence: 4 givenname: C.F. surname: Tu fullname: Tu, C.F. organization: Division of Biotechnology, Animal Technology Institute, Taiwan, P.O. Box 23, Chunan 35099, Miaoli, Taiwan – sequence: 5 givenname: C.S. surname: Lin fullname: Lin, C.S. email: lincs@mail.nctu.edu.tw organization: Department of Biological Science and Technology, National Chiao Tung University, No. 75 Po-Ai Street, Hsinchu 30068, Taiwan |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/18406177$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_1038_s41598_020_76715_w crossref_primary_10_1007_s00726_016_2249_5 crossref_primary_10_1080_02648725_2018_1506898 crossref_primary_10_1139_bcb_2013_0031 |
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| Keywords | Hi SI-PMEC pGB562/Hi EGFP pGB562/GFP/SI-PMEC Hirudin GB562 RT-PCR pGB562/Hi/SI-PMEC pGB562/GFP Epithelial cell Recombinant protein Mammary gland |
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| Snippet | To express recombinant proteins in the spontaneously immortalized porcine mammary epithelial cell line (SI-PMEC) currently established in our laboratory, a... To express recombinant proteins in the spontaneously immortalized porcine mammary epithelial cell line (SI‐PMEC) currently established in our laboratory, a... Abstract To express recombinant proteins in the spontaneously immortalized porcine mammary epithelial cell line (SI‐PMEC) currently established in our... |
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| SubjectTerms | Amino Acid Sequence Animals Base Sequence Cell Line Collagen - pharmacology Drug Combinations Epithelial cell Epithelial Cells - cytology Epithelial Cells - metabolism Fibrinolytic Agents - metabolism Gene Expression Genetic Vectors Hirudin Hirudins - biosynthesis Hirudins - genetics Laminin - pharmacology Mammary gland Mammary Glands, Animal Molecular Sequence Data Prolactin - pharmacology Proteoglycans - pharmacology Recombinant protein Recombinant Proteins - biosynthesis Recombinant Proteins - genetics SI-PMEC Swine Transfection |
| Title | Expression of recombinant anticoagulant hirudin in the differentiated cultures of the porcine mammary epithelial cell line SI-PMEC |
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