Wet lab works are emphasizing on the expression and purification of a variety of chimeric chitinases and determining their potency through a multitude of antifungal assays. The wet lab also dealt with learning basic lab techniques, the isolation of fungal samples from the campus, confirming the species identities, obtaining pure cultures of the fungal species and performing sanger sequencing using species-specific primers for identifying the fungal species and making pure cultures of the fungal species identified.
Chitinases are proteins that cleave the glycosidic bonds present in chitin, a major component of the cell walls of fungi. This can lead to cell lysis and resultant mortality. So, utilising the elegance of domain fusion and protein engineering, we aim to create novel chitinases with the potential to show enhanced antifungal activity.
Workflow of wet lab contained mainly the following steps:
- Designing chitinase-specific primers.
- Cloning of the chimeric chitinase genes
- Transformation of BL21 DE3 cells with the cloned plasmids.
- Induction of protein expression.
- Purification of the chitinases.
- Performing antifungal assays to measure the activity.