Establishment of an insect cell clone that harbours a partial baculoviral genome and is resistant to homologous virus infection.
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 1.25 x 10(5) 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.
Weng
Qingbei
Q
School of Life Sciences, Guizhou Normal University, Guiyang 550001, PR China.|State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou 510275, PR China.
Yang
Kai
K
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou 510275, PR China.
Xiao
Wei
W
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou 510275, PR China.
Yuan
Meijin
M
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou 510275, PR China.
Zhang
Wenjun
W
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou 510275, PR China.
Pang
Yi
Y
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou 510275, PR China.
D000818:Animals; D002460:Cell Line; D002999:Clone Cells; D016679:Genome, Viral; D017924:Nucleopolyhedroviruses / Q000145:classification / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D000077183:Occlusion Body Matrix Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012692:Serial Passage; D018411:Spodoptera / Q000821:virology*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism; D017735:Virus Latency
D000077183:Occlusion Body Matrix Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus