Abstract:
:Human interferon α (IFN-α) and erythropoietin (EPO) have been used for a variety of purposes in clinical medicine. Human IFN-α has been used to treat several types of viral infection and cancer, as well as renal anemia, via stimulation of erythrocyte formation in the bone marrow. Transgenic cattle are excellent candidates for pharmaceutical production for humans due to their ability to produce recombinant proteins in milk. The purpose of the present study was to generate bovine transgenic fibroblasts capable of producing recombinant human IFN-α and EPO proteins in transgenic cattle milk. First, we analyzed the promoter activities of various bovine milk protein genes in HC11 mouse mammary epithelial cells. The bovine milk protein gene promoters were cloned into the Luc gene in a promoter-less pGL3-Basic vector. Presence of the αS1-casein promoter (-175 to +796 nt) resulted in an up to 16-fold increase in luciferase activity compared with that of the promoter-less construct. In addition, the human IFN-α and EPO genes were identified as significantly overexpressed in HC11 cells compared with the promoter-less construct. Together, the present results demonstrate that the construct with the αS1-casein promoter may induce secretion of recombinant human IFN-α and EPO into bovine milk. Furthermore, we generated transgenic fibroblasts expressing human IFN-α and EPO cDNA controlled by the αS1-casein promoter and two screening markers, enhanced green fluorescent protein and neomycin resistance. These transgenic fibroblasts may be a source of somatic cells for generating transgenic cattle that produce recombinant human IFN-α and EPO proteins during lactation.
journal_name
Mol Med Repjournal_title
Molecular medicine reportsauthors
Jung EM,An BS,Kim YK,Hwang I,Lee JY,Shin TY,Hyun SH,Hwang WS,Jeung EBdoi
10.3892/mmr.2012.1182subject
Has Abstractpub_date
2013-02-01 00:00:00pages
406-12issue
2eissn
1791-2997issn
1791-3004journal_volume
7pub_type
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