Frank Nelissen

Stable isotope labeling of biological macromolecules for NMR

My work mainly focuses on the enzymatic synthesis of homogeneous and highly pure biological macromolecules such as DNA and RNA in amounts sufficient for NMR applications. To enhance the sensitivity of these biomolecules for NMR, labeling with stable isotopes (Carbon-13 and Nitrogen-15) is required. The labeling pattern of a biomolecule can either be uniformly, (multiple) segmental, residue-type selective or residue selective. In addition, (stereo-)selective deuteration or perdeuteration can be applied to reduce resonance overlap and line widths in NMR spectra.

In order to be able to produce such specifically labeled samples, new and existing (bio)chemical, molecular biological and analytical techniques are developed and used in combination. Tailor labeled stable isotope labeled building blocks (rNTPs, dNTPS) and the majority of the required production enzymes are in-house produced and purified. Furthermore, stable isotope labeled metabolites with cellular regulatory functions (e.g. S-adenosylmethionine) can be synthesized enzymatically, dependent on the research question.

Next generation sequencing

Using massive parallel sequencing, RNA elements and RNA sequences involved in gene expression regulation in living organisms are sought (e.g. frame-shifting sequences in yeast). Without the use of commercial sample work-up kits, I am able to prepare the required sequencing libraries according to established protocols that yield whole transcriptome information (RNA-Seq), translational information (Ribo-Seq) and structure-dependent RNA function information (Structure-Seq).

Support

I spend considerable time of my work in assisting Postdocs, PhD students and master/bachelor students of our own group as well as from affiliated groups in their research. This varies between conducting whole experiments, providing advice for experimental setup, giving instructions for the use of equipment and ordering chemicals and consumables.

Other activities

I am the First Aid person for our department as well as the Health & Safety Officer concerning working circumstances and therefore involved by everything that goes on in these fields. Furthermore I am the Responsible Employee for the registration and observation of everything concerning Genetically Modified Organisms in our ML-1 laboratory.