We describe the use of herpesvirus promoters to regulate the expression of a @entity12385 replicon (SINrep/LacZ). We isolated cell lines that contain the cDNA of SINrep/LacZ under the control of a promoter from a herpesvirus early gene which requires regulatory proteins encoded by immediate-early genes for expression. Wild-type @entity12385 and replicons derived from this virus cause death of most vertebrate cells, but the cells discussed here grew normally and expressed the replicon and beta-galactosidase only after infection with a herpesvirus. Vero cell lines in which the expression of SINrep/LacZ was regulated by the @entity3307 ) infected-cell protein 8 promoter were generated. One Vero cell line (V3-45N) contained, in addition to the SINrep/LacZ cDNA, a @entity12385 -defective helper cDNA which provides the structural proteins for packaging the replicon. Infection of V3-45N cells with @entity3307 resulted in the production of packaged SINrep/LacZ replicons. @entity3307 induction of the @entity12385 replicon and packaging and spread of the replicon led to enhanced expression of the reporter gene, suggesting that this type of cell could be used to develop sensitive assays to detect herpesviruses. We also isolated a mink lung cell line that was transformed with SINrep/LacZ cDNA under the control of the promoter from the @entity4538 ) early gene @entity68793 . HCMV carries out an @entity281 in mink lung cells, but it was able to induce the SINrep/LacZ replicon. These results, and those obtained with an @entity3307 mutant, demonstrate that this type of signal amplification system could be valuable for detecting herpesviruses for which a permissive cell culture system is not available.

Regulated expression of a XXXX replicon by herpesvirus promoters.

@entity4538 :: ('10359', 'Species') :: ['human cytomegalovirus (HCMV'] @entity281 :: ('MESH:D007239', 'Disease') :: ['abortive infection'] @entity3307 :: ('10298', 'Species') :: ['herpes simplex virus type 1 (HSV-1', 'HSV-1'] @entity12385 :: ('11034', 'Species') :: ['Sindbis virus'] @entity68793 :: ('3077444(Tax:10359)', 'Gene') :: ['UL45']
@entity12385 :: (11034,Species) :: ['Sindbis virus']
