1.Sequence analysis of polyhedrin gene promoter and construction of an expression vector of hyphantria cunea nuclear polyhedrosis virus.
Kap Joo PARK ; Bong Joo KANG ; Hye Kyung CHUNG ; Bon Hong MIN ; Hyung Hoan LEE
Journal of the Korean Society of Virology 1993;23(2):141-151
No abstract available.
Nucleopolyhedrovirus*
;
Sequence Analysis*
2.Location and cloning of the polyhedrin gene of hyphantria cunea nuclear polyhedrosis virus.
Hyung Hoan LEE ; Mi Kyung LEE ; Il Hwan CHO ; Kwan Hee YOO
Journal of the Korean Society of Virology 1991;21(1):25-34
No abstract available.
Clone Cells*
;
Cloning, Organism*
;
Nucleopolyhedrovirus*
4.Characterization of Insect Cells Transformed with Autographa calfornica Nuclear Polyhedrosis Virus IE1 Gene.
Eun Sook CHO ; Hae Jin PARK ; Kwang Sik LEE ; Seok Woo KANG ; Eun Young YUN ; Keun Young KIM ; Hung Dae SOHN ; Byung Rae JIN
Journal of the Korean Society of Virology 1999;29(2):137-144
Transformation efficiency, virus multiplication and foreign gene expression were characterized in the insect cells transformed with Autographa calfornica nuclear polyhedrosis virus (AcNPV) immediate early 1 gene (IE1). Transformation efficiency of insect cells by AcNPV IE1 gene vector horboring foreign gene was approximately 8-fold higher in the Sf9 cells transformed previously with AcNPV IE1 gene than in the normal Sf9 cells. Virus multiplication and foreign gene expression of recombinant baculovirus in the Sf9 cells transformed with AcNPV IE1 gene were similar to those of the normal Sf9 cells. These results suggest that transformed cells displaying foreign gene product by using AcNPV IE1 gene promoter will be useful for the diverse applications of insect cells.
Baculoviridae
;
Gene Expression
;
Insects*
;
Nucleopolyhedrovirus*
;
Sf9 Cells
5.Characterization of Insect Cells Transformed with Autographa calfornica Nuclear Polyhedrosis Virus IE1 Gene.
Eun Sook CHO ; Hae Jin PARK ; Kwang Sik LEE ; Seok Woo KANG ; Eun Young YUN ; Keun Young KIM ; Hung Dae SOHN ; Byung Rae JIN
Journal of the Korean Society of Virology 1999;29(2):137-144
Transformation efficiency, virus multiplication and foreign gene expression were characterized in the insect cells transformed with Autographa calfornica nuclear polyhedrosis virus (AcNPV) immediate early 1 gene (IE1). Transformation efficiency of insect cells by AcNPV IE1 gene vector horboring foreign gene was approximately 8-fold higher in the Sf9 cells transformed previously with AcNPV IE1 gene than in the normal Sf9 cells. Virus multiplication and foreign gene expression of recombinant baculovirus in the Sf9 cells transformed with AcNPV IE1 gene were similar to those of the normal Sf9 cells. These results suggest that transformed cells displaying foreign gene product by using AcNPV IE1 gene promoter will be useful for the diverse applications of insect cells.
Baculoviridae
;
Gene Expression
;
Insects*
;
Nucleopolyhedrovirus*
;
Sf9 Cells
6.Construction of a Novel Recombinant Baculovirus Producing Polyhedra with Bacillus thuringiensis Cry1Ac Crystal Protein.
Yeon Ho JE ; Byung Rae JIN ; Jong Yul ROH ; Jin Hee CHAN ; Seok Kwon KANG
Journal of the Korean Society of Virology 1999;29(3):145-153
We have now construted a novel recombinant baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) producing polyhedra with Bacillus thuringiensis (Bt) Cry1Ac crystal protein. The recombinant polyhedra produced by the recombinant baculovirus, Btrus, in insect cells was characterized. The recombinant baculovirus has two independent transcription units in opposite orientations with two promoters, p10 or polyhedrin gene promoter each initiating transcription of either native polyhedrin or fusion protein with polyhedrin and Bt Cry1Ac crystal protein. Suprisingly, this recombinant baculovirus stably produced recombinant polyhedra which were nearly similar to those of wild-type AcNPV. The immunogold staining experiment showed that the recombinant polyhedra were assembled with polyhedrin and Bt Cry1Ac crystal protein, and contained virus paticles. Insecticidal toxicity of recombinant polyhedra of Btrus to the fall webworm, Hyphantrea cunea, was strikingly improved in comparison with the wild-type AcNPV.
Bacillus thuringiensis*
;
Bacillus*
;
Baculoviridae*
;
Insects
;
Nucleopolyhedrovirus
7.Cloning of the hyphantrica cunea nuclear polyhedrosis virus partial EcoRI genome DNA fragments in plasmid vectors pUC8 and pBR322.
Hyung Hoan LEE ; Jin Wook KIM ; Hee Kyung KIM ; Sung Sook PARK ; Yong Chull LEE ; Dong Chull OK
Journal of the Korean Society of Virology 1991;21(1):35-40
No abstract available.
Clone Cells*
;
Cloning, Organism*
;
DNA*
;
Genome*
;
Nucleopolyhedrovirus*
;
Plasmids*
8.Characteristics and Pathogenicity of Host Range Expanded Recombinant Viruses in Insect Cells.
Hye Sung KIM ; Soo Dong WOO ; Woo Jin KIM ; Jae Young CHOI ; Byung Rae JIN ; Youn Hyung LEE ; Seok Kwon KANG
Journal of the Korean Society of Virology 1997;27(1):29-37
To use recombinant viruses with wider host range as viral insecticides, we investigated the characteristics and pathogenicity of host range expanded recombinant viruses in insect cells. We compared host range expanded recombinant viruses, RecS-B6 and RecB-8, constructed by cotransfection of Autographa californica nuclear polyhedrosis virus (AcNPV) and Bombyx mori NPV (BmNPV), to host range expanded AcNPV, Ac-BH, by substitution of the 0.6 Kb fragment of the BmNPV helicase gene. Restriction endonuclease profiles of RecS-B6 and RecB-8 DNAs were different from those of parent viruses. Nucleotide sequence analysis of the 0.6 Kb region in the putative helicase gene of RecS-B6 and RecB-8 showed that their structures were identical to the counterpart region of BmNPV Comparison of viral replication of these recombinant viruses in Sf-21 and BmN-4 cells showed that Ac-BH, compared to wild type viruses, replicated well in BmN-4 cells but poorly in Sf-21 cells. In contrast, RecS-B6 and RecB-8 replicated relatively well in both cells compared to parent viruses. These results may imply that random genomic recombinant viruses, RecS-B6 and RecB-8, possess better potential as viral pesticides than helicase-mediated recombinant virus, Ac-BH.
Base Sequence
;
Bombyx
;
DNA
;
DNA Restriction Enzymes
;
Host Specificity*
;
Humans
;
Insecticides
;
Insects*
;
Nucleopolyhedrovirus
;
Parents
;
Pesticides
;
Virulence*
9.AcMNPV-mediated expression of BmK IT promotes the apoptosis of Sf9 cells and replication of AcMNPV.
Yue-Jun FU ; Jie ZHAO ; Ai-Hua LIANG ; Feng-Yun HU
Acta Physiologica Sinica 2015;67(3):305-311
Chinese scorpion Buthus martensii Karsch (BmK) venom is a rich source of neurotoxins which bind to various ion channels with high affinity and specificity and thus widely used as compounds to modulate channel gating or channel currents. To promote the insecticidal effects of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), the gene encoding an excitatory insect toxin, BmK IT, was inserted into the genome of AcMNPV to construct a recombinant baculovirus, AcMNPV-BmK IT. Spodopter frugiperda 9 (Sf9) cells were infected with AcMNPV and AcMNPV-BmK IT respectively for 24 h. Results from the MTT assay, TUNEL assay, analysis of the expression level of apoptosis-related proteins (c-Myc, cleaved-Caspase3, Bcl-2 and Bax) of Sf9 cells, the transcription level of key genes (38K, C42, P78, F) of AcMNPV, and viral propagation assay demonstrated that AcMNPV-mediated expression of BmK IT promoted the apoptosis of Sf9 cells and replication of AcMNPV. The results laid a foundation for further structural and functional analysis of BmK IT.
Animals
;
Apoptosis
;
Cell Line
;
Nucleopolyhedrovirus
;
metabolism
;
physiology
;
Scorpion Venoms
;
biosynthesis
;
Sf9 Cells
;
drug effects
;
Virus Replication
10.Comparative study of the replication difference of HearNPV in infected exponential and stationary host cells.
Wen-Tao DAI ; Xiao HAN ; Hua-Lin WANG ; Zhi-Hong HU ; Fei DENG
Chinese Journal of Virology 2007;23(5):399-406
Real-time quantitative PCR was used to characterize HearNPV DNA replication in exponential and stationary phases of HzAM1 cells. Results showed that the doubling time of HzAM1 cells was 22 h in exponential phases. Most of the exponential cells were in S phase (48.6%), and most of the stationary cells in G2/M phase (72.6%). The replication of viral DNA was completed within 60 h post infection (h p. i.) in different phases of HzAM1 cells. During 14 to 20 h p. i., the doubling time of HearNPV replica-tion was 1.8 h in exponential cells and 1.9 h in stationary cells, and no significant difference was found between them. But the amounts of BV entering and releasing, the final progeny virions and viral protein products in the infected exponential phase cells were obviously higher than that in the stationary phase cells. 25% of the total synthesized viral DNAs were released from infected exponential phase cells, but on-ly 13% from the infected stationary phase cells. Viral DNA started to be replicated from 7-8 h p. i. both in infected exponential phase and in stationary phase cells. But in infected exponential phase cells, BVs were started to release from 18-20 h p. i., and BVs were started to release from 22-25 h p. i. from infected sta-tionary phase cells. During 30-60 h p. i., the BV releasing rate was about 483 copies/cell/h in the expo-nential phase cells, but was 100 copies/cell/h in the stationary-phase cells. The initial viral DNA entering into exponential phase cells was much more than that entered into the stationary phase cells. The data of cell membrane fluidity at exponential and stationary phases suggested that the fluidity of cell membrane played an important role during virus entry.
Animals
;
Cell Cycle
;
Cell Line
;
DNA Replication
;
Membrane Fluidity
;
Moths
;
Nucleopolyhedrovirus
;
physiology
;
Virus Internalization
;
Virus Replication