Construction and characterization of a Helicoverpa armigera nucleopolyhedrovirus bacterial artificial chromosome with deletion of ecdysteroid UDP-glucosyltransferase gene.
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.
Si
Yan-Hong
YH
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, PR China.
Fang
Ming-Gang
MG
Huang
Yi
Y
Zheng
Fang-Liang
FL
Li
Ting
T
Hu
Zhi-Hong
ZH
Wang
Han-Zhong
HZ
D000818:Animals; D001681:Biological Assay; D022202:Chromosomes, Artificial, Bacterial; D017353:Gene Deletion; D005814:Genes, Viral; D005818:Genetic Engineering; D005964:Glucosyltransferases / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007814:Larva / Q000648:ultrastructure / Q000821:virology; D007928:Lethal Dose 50; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011995:Recombination, Genetic; D015183:Restriction Mapping; D013997:Time Factors
D049452:Green Fluorescent Proteins; D005964:Glucosyltransferases; C086659:ecdysteroid UDP-glucosyltransferase