Analysis of promoter activity of selected Cotesia plutellae bracovirus genes

Abstract

In a previous study, we cloned 27 discrete genome segments of Cotesia plutellae bracovirus (CpBV) and provided the complete nucleotide sequences and annotation. Seven putative coding regions were predicted from one of the largest segments, CpBV-S30. The activity of promoters associated with six predicted ORFs from this segment were investigated using both transient and baculovirus expression assays with enhanced green fluorescent protein as a reporter gene. CpBV promoters showed activity earlier than the polyhedrin promoter and the activity of some of these promoters was superior to that of the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ie-1 promoter in the baculovirus expression assays. The promoter of ORF3004 showed the highest level of activity in insect cells, exhibiting 24 % of the activity obtained with the AcMNPV polyhedrin promoter in Sf9 cells. In Spodoptera exigua larvae, the ORF3006 promoter showed the highest activity, with about 35 % of the activity measured with the polyhedrin promoter. In addition, analysis of the ORF3006 promoter revealed that the region between "382 and "422 from the translation start point was critical for activity of this promoter. These results suggest that the CpBV-S30 promoters characterized here could be useful tools in a variety of biotechnological applications, such as gene expression analyses and insecticide development.

NeuroBactrus, a Novel, Highly Effective, and Environmentally Friendly Recombinant Baculovirus Insecticide

“…The CpBV ORF3006 promoter used for expression of AaIT in this recombinant virus showed activity as strong as ca. 35% of that of the polyhedrin promoter in the hemolymphs of S. exigua larvae (30). The fused Cry1-5 toxin could be activated in the midgut of host insects through proteolytic cleavage of the toxin by proteases.…”

A novel polydnaviral gene family, BEN, and its immunosuppressive function in larvae of Plutella xylostella parasitized by Cotesia plutellae

“…Vankyrin and EP gene families are also found in CpBV each with six members (Kwon and Kim, 2008;Bae and Kim, 2009). Except for BEN family, other known CpBV gene families are determined in their expression in parasitized host and physiological functions in the parasitism Choi et al, 2009). To propose 11 CpBV-BEN genes as a novel functional PDV gene family, this study analyzed their gene expressions in larvae of P. xylostella parasitized by C. plutellae.…”

Bombyx mori Nucleopolyhedrovirus Bacmid Enabling Rapid Generation of Recombinant Virus by In Vitro Transposition

“…Barnase, a major extracellular ribonuclease isolated from B. amyloliquefaciens, is lethal to cells in which it is expressed [5,15]. The CpBV promoter used for the expression of barnase showed activity earlier than the polyhedrin promoter, and the activity of some of these promoters was superior to the AcMNPV ie-1 promoter [3]. The bEasyBm can only replicate in transfected insect cells when the CpBV promoterbarnase cassette is replaced with the gene of interest by in vitro transposition.…”