MOS55 cells were infected with recombinant DNV sample corresponding to 5 × 108 of recombinant virus. For quantification of the viral DNA genome, the supernatant of infected cells was collected at indicated days after infection. EGFP expression in the cell line was observed using an Axiovert S100 fluorescence microscope (Zeiss) and images captured using a ProgRes CF camera (Jenoptik). EGFP signal was quantified by flow cytometry on a Cytomics FC500 (Beckman Coulter). Data were analyzed using BD LSRFortessa (BD Biosciences) and FlowJo software. Anopheles gambiae adults were intrathoracically injected according to Hughes et al. (2011)68 (link) with the indicated viral copy number of recombinant DNV virions. Fluorescence in mosquitoes and dissected tissues was monitored at indicated days post-injection using an Olympus BX-41 microscope. Images were processed using PictureFrame software (Olympus).
Protocol detail
Quantification of Recombinant DNV Infection
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Adults
Anopheles gambiae
Cell line
Flow cytometry
Fluorescence
Fluorescence microscope
Genome
Infection
Microscope
Mosquitoes
S100
Tissues
Viral dna
Viral genome
Virions
Virus
Corresponding Organization :
Other organizations : Pennsylvania State University
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Variable analysis
independent variables
- Infection of MOS55 cells with recombinant DNV sample corresponding to 5 × 10^8 of recombinant virus
- Intrathoracic injection of Anopheles gambiae adults with the indicated viral copy number of recombinant DNV virions
- Quantification of the viral DNA genome in the supernatant of infected cells at indicated days after infection
- EGFP expression in the cell line observed using an Axiovert S100 fluorescence microscope and quantified by flow cytometry on a Cytomics FC500
- Fluorescence in mosquitoes and dissected tissues monitored at indicated days post-injection using an Olympus BX-41 microscope
- Not explicitly mentioned
- Positive control: Not mentioned
- Negative control: Not mentioned
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