Utilization of an Epstein-Barr virus replicon as a eukaryotic expression vector.
Epstein-Barr virus (EBV) replicons which include the genetic element oriP and a functional gene for Epstein-Barr nuclear antigen (EBNA-1) can be maintained episomally in a variety of mammalian cell lines [Yates et al., Nature 313 (1985) 812-815]. We have assessed the application of an EBV replicon for foreign gene expression. Two cDNAs, human interferon-gamma (IFN-gamma) and the extracellular domain of the human epidermal growth factor receptor (EGF-Rex), cloned in an EBV replicon, were efficiently expressed and the protein was secreted into the extracellular media. Expression in human embryonic 293 cells was approximately ten-fold higher than in CV-1 cells. The expression of the human protein is dependent upon the orientation of the IFN-gamma transcriptional cassette relative to the other genetic elements within the vector.
Young
J M
JM
Molecular Biology Division, Rorer Biotechnology, Inc., Springfield, VA 22151.
Cheadle
C
C
Foulke
J S
JS
Drohan
W N
WN
Sarver
N
N
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D002460:Cell Line; D002477:Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods*; D004274:DNA, Recombinant / Q000032:analysis; D004622:Embryo, Mammalian; D019309:Epstein-Barr Virus Nuclear Antigens; D066246:ErbB Receptors / Q000096:biosynthesis; D005057:Eukaryotic Cells; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics; D007668:Kidney; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012093:Replicon
D000956:Antigens, Viral; D004274:DNA, Recombinant; D019309:Epstein-Barr Virus Nuclear Antigens; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D007371:Interferon-gamma; D066246:ErbB Receptors