PROPAGATION AND RECOVERY OF INTACT, INFECTIOUS EPSTEIN-BARR-VIRUS FROM PROKARYOTIC TO HUMAN-CELLS

With current techniques, genetic alterations of herpesviruses are diff icult to perform, mostly because of the large size of their genomes, T o solve this problem, we have designed a system that allows the clonin g of any gamma-herpesvirus in Escherichia coli onto an F factor-derive d plasmid, Immortalized B cell lines were readily established with rec ombinant Epstein-Barr virus (EBV), demonstrating that the F factor-clo ned EBV genome has all the characteristics of wild-type EBV, Because a ny genetic modification is possible in E, coli, this experimental appr oach opens the way to the genetic analysis of all EBV functions, Moreo ver, it is now feasible to generate attenuated EBV strains in vitro su ch that vaccine strains can be designed. Because we incorporated the g enes for hygromycin resistance and green fluorescent protein onto the E. coil cloned EBV genome, the still open question of the EBV target c ells other than B lymphocytes will be addressed.