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Establishment of an insect cell clone that harbours a partial baculoviral genome and is resistant to homologous virus infection

¹ State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou 510275, PR China
² School of Life Sciences, Guizhou Normal University, Guiyang 550001, PR China

Correspondence
Yi Pang
pangy{at}mail.sysu.edu.cn

After serially undiluted passage of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV), persistently infected Se301 cells were established. A cell strain, in which no polyhedra or viral particles were observed, was cloned and designated P8-Se301-C1. The P8-Se301-C1 cells are morphologically similar to but grow slower than Se301 cells and they can homologously interfere with SeMNPV. PCR analysis showed that SeMNPV ie-0 and polyhedrin genes were present but DNA polymerase and orf67 genes were absent in P8-Se301-C1, suggesting that the cells harbour incomplete SeMNPV genomes. Dot-blot analysis demonstrated that 0.32±0.16 ng SeMNPV DNA was present in 1.25x10⁵ P8-Se301-C1 cells. A quantitative real-time PCR assay showed that there were 13.2±4.3 copies of the SeMNPV polyhedrin gene in each cell. Nested RT-PCR demonstrated the presence of SeMNPV polyhedrin transcripts in P8-Se301-C1 cells. The fact that P8-Se301-C1 cells carry low levels of partial viral genome but do not produce viral progeny suggests a latent-like viral infection in the cells.