Molecular integrity and usefulness of episomal expression vectors derived from BK and Epstein-Barr virus.
High-level and stable production of a protein of interest is one of the most important parameters when considering the development of an efficient vector system for heterologous gene expression. In order to achieve this goal, we have used episomal vector elements derived from Epstein-Barr virus (EBV) or BK virus (BKV) in combination with the strictly regulated interferon-inducible Mx promoter. Here we demonstrate that EBV-derived vectors replicate efficiently in all cell lines tested (i.e. HEK293, HeLaH21 and Vero), yielding stable transfectants with a high, inducible expression level and almost no background. In contrast, BKV-derived vectors are much more restricted to particular cell types and hampered by DNA rearrangements, which is a serious drawback for use over a longer timespan.
Department of Molecular Biology, University of Gent-VIB, K.L. Ledeganckstraat 35, B-9000, Gent, Belgium.
K
K
Van Craenenbroeck
P
P
Vanhoenacker
H
H
Duchau
G
G
Haegeman
D000818:Animals; D001739:BK Virus / Q000235:genetics*; D002460:Cell Line; D002522:Chlorocebus aethiops; D004261:DNA Replication / Q000235:genetics; D004274:DNA, Recombinant; D005057:Eukaryotic Cells / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics*; D006367:HeLa Cells; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D014162:Transfection; D014709:Vero Cells
D004274:DNA, Recombinant