Construction and Characterization of a Helicoverpa armigera Nucleopolyhedrovirus Bacterial Artificial Chromosome with Deletion of Ecdysteroid UDP-Glucosyltransferase Gene

Abstract

Genetically modified baculoviruses offer a promising alternative to chemical insecticides in the control of agricultural and forest insect pests. A novel bacmid, HaBacYH6, was constructed in which the ecdysteroid UDP-glucosyltransferase gene (egt) was replaced with a bacterial replication cassette containing a mini-F replicon, a kanamycin resistance gene, and the attTn7 site. Insertion of the enhanced green fluorescence protein gene (egfp) into HaBacYH6 showed that the bacmid can express an active exogenous protein. Bioassays showed that median lethal time (LT₅₀) of HaBacYH6 was 89.23 h in third instar Helicoverpa armigera larvae, 15.81 h earlier than that of wild-type HearNPV-G4, though there was no significant difference in median lethal dose (LD₅₀). The data indicate that HaBacYH6 can be used as a new Bac-to-Bac system, and can also provide a technology platform for generating more effective biological insecticides.