Ready for take off: Set up and start of nudivirus experiments
November 8, 2021
My name is Jirka Petersen and I am affiliated with the INSECT DOCTORS program as ESR1. I moved from Germany to Wageningen (Netherlands) last year in August. Since then I’ve been setting up the laboratory for my experiments with the Heliothis zea nudivirus 1 (HzNV-1). Interestingly, this lepidopteran nudivirus can only be maintained in cell culture and is unable to infect actual insects.
Therefore, the first months of my laboratory work involved the establishment of a HzNV-1 production system. In the process we were dealing with a baculovirus contamination in the original HzNV-1 isolate that we were able to purify away through several passaging in baculovirus-resistant HzAM1 cells. With the pure HzNV-1 stock we were then able to conduct follow-up experiments in these cells, such as bioassays, (electron) microscopic analyses, DNA and RNA extractions, and cloning approaches.
One experiment aims to study the viral and host gene expression profiles during infection of HzAM1 cells with HzNV-1 and is now in progress. Another assay is being developed to capture viral proteins involved in HzNV-1 replication. Furthermore, degenerate primers for the detection of different nudivirus species were designed for the development of an infection diagnostic tool in insect rearing. At this stage, the conditions for the degenerate primers are still being optimized using HzNV-1 as a template, before their application on actual insect samples.
During a training on how to operate the electron microscope and preparation of samples in suspension and tissue, we obtained yet undescribed insight in the cell entry mechanism of HzNV-1. These results provide new experimental leads on how to study the cytopathogenicity of this nudivirus.