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SPP1 bacteriophage infects the Gram-positive model bacterium Bacillus subtilis and is a well-characterized biological system. Recent studies from our laboratory have shown that during infection SPP1 forms a spatially independent factory for viral genome replication in the bacterial cytoplasm. In my current project, I aim to characterize the molecular and cellular mechanisms underlying the formation of such viral replication factories using far-end fluorescence microscopy methods. The dynamics of the phage DNA and replication proteins will be imaged using single particle tracking (SPT) and microfluidics to follow the complete infection cycle in real time. Phage mutants will be used to arrest assembly of the replication factory at specific steps to determine the order of recruitment of their individual components and to study transient states of the replication factory. The organization and dynamics of the factories will be imaged employing single-molecule tracking (SMT). Finally, the recruitment of bacterial replisome proteins to the viral genome replication factory will also be studied. The ultimate aim of this work is to reveal the underlying mechanisms used by viruses to invade bacterial cells and convert them into virus factories.

1. F. Mayer et al. Luminescence properties of self-aggregating TbIII-DOTA-functionalized Calix[4]arenes. Front. Chem. (2018), doi: 10.3389/fchem.2018.00001
2. S. Tiruvadi Krishnan et al. The progression of replication forks at natural replication barriers in live bacteria. Nucleic Acids Res, (2016), 44 (13): 6262-6273. doi 10.1093/nar/gkw397
3. S. Tiruvadi Krishnan et al. Essential validation methods for Escherichia coli strains created by chromosome engineering. J.Biol.Engg (2015), 9:11. DOI 10.1186/s13036-015-0008-x
4. M. C. Moolman et al. Slow unloading leads to DNA-bound β2 sliding clamp accumulation in live Escherichia coli cells. Nat.Comm (2014), 5:5820. DOI: 10.1038/ncomms6820
5. M. C. Moolman et al. Electron beam fabrication of a microfluidics device for submicron-scale bacteria. J.Nanobiotechnology (2013), 11:12. doi:10.1186/1477-3155-11-12

09/2010 – 09/2016
Ph.D. in Cellular biophysics at Nynke Dekker lab, Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology (TUDelft), The Netherlands.

Specialization: Genome engineering & fluorescence microscopy.

Thesis title: In vivo investigations of E. coli chromosomal replication using single-molecule imaging.

09/2008 - 08/2010
Master in Life science and technology at TUDelft, The Netherlands.

Specialization: Cell diagnostics & molecular biology.

Thesis title: Developing metalloorganic contrast agents for early MRI diagnosis of tumor and targeted drug delivery.