Development of a novel expression vector system using Spodoptera exigua nucleopolyhedrovirus.
To develop a novel Spodoptera exigua nucleopolyhedrovirus (SeNPV) expression vector system, we examined characteristics of the SeNPV polyhedrin expression in S. exigua cells (Se301). While the extracellular virus titer of SeNPV was 100-fold lower than that of Autographa californica nucleopolyhedrovirus (AcNPV), the levels of polyhedral inclusion body (PIB) formation and polyhedrin expression were higher in SeNPV. To investigate foreign gene expression under the control of the polyhedrin promoter, polyhedrin-based transfer vector pSeKSK2 was constructed, and then recombinant virus SeK1-LacZ was constructed by inserting E. coli lacZ gene as a reporter gene into a genomic DNA of SeNPV using this transfer vector. The beta-Galactosidase activity of SeK1-LacZ in Se301 was about 1.3 times higher than that of BacPAK6. Thus, the SeNPV expression vector system constructed in this study would be very useful in the expression of foreign proteins, specifically for the enhancement of the pesticidal properties of SeNPV by inserting pesticidal genes.
Choi
J Y
JY
Division of Applied Biology and Chemistry, College of Agriculture and Life Sciences, Seoul National University, Suwon, Korea.
Woo
S D
SD
Je
Y H
YH
Kang
S K
SK
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D007763:Lac Operon; D009036:Moths / Q000166:cytology / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D000077183:Occlusion Body Matrix Proteins; D011401:Promoter Regions, Genetic; D013045:Species Specificity; D018411:Spodoptera / Q000166:cytology / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins; D014779:Virus Replication; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D000077183:Occlusion Body Matrix Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D001616:beta-Galactosidase