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Updated: Aug 9, 2026
Transient Expression and Cellular Localization of Recombinant Proteins in Cultured Insect Cells
Published on: April 20, 2017
Bacterial expression and cellular localization of Helicoverpa armigera nucleopolyhedrovirus Orf33 in infected host
Dun Wang1, Xing-cheng Yan, Kumar Shyam
1Institute of Applied Entomology, Hangzhou 310029, China.
Abstract:
Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearSNPV) is a selective, highly infectious pathogen to H. armigera and has been extensively used for the control of this pest. This study presents the bacterial expression and localization of ha33 in infected host cells. The ha33 protein expressed in Escherichia coli had protein size of 17kDa. Western blot analysis using polyclonal antibody showed that the product of ha33 in infected Helicoverpa zea cells (HzAM1) was a 31kDa protein, larger than the theoretically predicted size of 28.4kDa. The confocal laser scanning microscope observation demonstrated that ha33 gene product was localized in the cytoplasm of infected HzAM1 cells beginning at 6 h p. i. and remained throughout the infection.
Insights
Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearSNPV) protein ha33 was expressed in E. coli and its localization in infected cells was studied. The ha33 protein was found in the cytoplasm of infected HzAM1 cells.
Area of Science:
- Virology
- Molecular Biology
- Insect Pathology
Background:
- Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearSNPV) is a vital biocontrol agent for Helicoverpa armigera.
- Understanding viral protein function and localization is crucial for optimizing biocontrol strategies.
Purpose of the Study:
- To investigate the bacterial expression and cellular localization of the ha33 protein from HearSNPV.
- To determine the size and location of the ha33 protein within infected host cells.
Main Methods:
- Bacterial expression of ha33 in Escherichia coli.
- Western blot analysis using polyclonal antibodies to detect ha33 in infected Helicoverpa zea cells (HzAM1).
- Confocal laser scanning microscopy to visualize ha33 localization in infected HzAM1 cells.
Main Results:
- The ha33 protein expressed in E. coli showed a size of 17kDa.
- Western blot analysis revealed the ha33 protein in infected HzAM1 cells to be 31kDa, larger than its predicted size of 28.4kDa.
- Confocal microscopy showed ha33 localized in the cytoplasm of HzAM1 cells starting at 6 hours post-infection.
Conclusions:
- The ha33 protein is expressed and correctly localized in the cytoplasm of HearSNPV-infected host cells.
- Post-translational modifications may account for the larger-than-predicted size of the ha33 protein in infected cells.
- This study provides insights into the molecular mechanisms of HearSNPV infection and aids in developing improved biocontrol agents.
