SERIAL PASSAGE OF TRICHOPLUSIA NI SINGLE EMBEDDED BACULOVIRUS IN HOMOLOGOUS CELL LINE*

Abstract

Trichoplusia ni single embedded nuclear polyhedrosis virus (TnSNPV) is highly pathogenic for cell line but there were some problems with producing a few polyhedra and decrease of efficiency during serial passages. Sixty serial passages of TnSNPV were conducted in Tn 5B1–4 cell. Replication of the viral DNA and comparison of the viral character between the wild virus and clones from serial passages were performed. The TnSNPV DNA molecular weight, 115.8 kbp, was estimated by restriction enzyme analysis. Replication of the viral DNA which was analyzed by slot blot hybridization started at 8 h postinfection (p.i) and the DNA increased fast after 28 h p. i. The DNA synthesis reached a maximal number at 40 h p. i. The result from serial passage of the virus demanstrated that the relative BV titer was maitained at approximately 5 log TCIDSO/mL. The polyhedra of viral clones from early passages were almost normal but the majority of clones from late passages produced polyhedra without virions by examination with electron microscopy. Although there were some alterations with viral DNA from different clones, yet all of clones were from a homologous genome of wild virus by examination of restriction enzyme analysis and DNA:DNA hybridization.