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An OriP/EBNA‐1‐based baculovirus vector with prolonged and enhanced transgene expression
Author(s) -
Shan Liang,
Wang Leyao,
Yin Juan,
Zhong Peng,
Zhong Jiang
Publication year - 2006
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.978
Subject(s) - autographa californica , biology , transgene , transduction (biophysics) , hek 293 cells , green fluorescent protein , gene , recombinant dna , cell culture , microbiology and biotechnology , genome , gene delivery , transfection , vector (molecular biology) , vero cell , gene expression , virology , virus , plasmid , genetics , biochemistry , spodoptera
Background The baculovirus Autographa californica multiple nucleopolyhedrovirus ( Ac MNPV) has been explored as a gene delivery vehicle for a variety of mammalian cell lines. However, the transient expression nature due to its incapability to replicate in mammalian cells and insufficient transduction efficiency limit its application. Methods Recombinant baculovirus vectors containing genetic elements from Epstein‐Barr virus (EBV), OriP and EBNA‐1 , which are essential for the episomal maintenance of the EBV genome in latently infected cells, were constructed and tested for their ability to sustain and express transgene (enhanced green fluorescence protein ( egfp )) in mammalian cells. Results The recombinant baculovirus containing OriP and EBNA‐1 genes driven by the cytomegalovirus (CMV) promoter was capable of persisting in a significant proportion of infected mammalian cells, HEK293, Vero, Cos‐7, and Hone‐1, without any selective pressure. In HEK293, the expression of EGFP lasted for 60 days with markedly enhanced expression level. The persistence of baculovirus genome correlated with the expression of EBNA‐1. Conclusions The improved baculovirus vector could mediate prolonged and enhanced foreign gene expression in some mammalian cells. Furthermore, an adequate level of the EBNA‐1 protein was essential for the maintenance of the OriP ‐containing baculovirus genome. The new vector has potential for use in gene therapy. Copyright © 2006 John Wiley & Sons, Ltd.