Congratulations Harsh!
On April 7th, Harsh Yadav defended his MSc thesis in Biology-Biotechnology. Harsh’s project was entitled " Generating and screening GTR2 transporter mutant libraries for detection of hyperactive mutants”.
From a biotechnological perspective transport processes represent attractive targets for modulation of metabolite levels and are the foundation for the emerging field of transport engineering. Potential applications of transport engineering include control of metabolite accumulation in a tissue-specific manner in crop plants as well as increased yields of commercially valuable compounds produced in synthetic biology approaches. To unleash the potential of transport engineering we not only need methods to identify target transporters but also methods to engineer their properties according to our needs. However, at the moment we lack basic insights into the molecular determinants of transport properties of the NPF family.
In his thesis, Harsh focused on developing GTR2 as a model system to study the structure-function relationship of specialized metabolite transporters belonging to the NPF family. He used targeted mutagenesis and automated two electrode voltage clamp electrophysiology to study a point mutation conferring hyperactivity to GTR2. Harsh also succeeded in expressing GTR2 in yeast and showed that the hyperactivity was detectable in yeast as well as in Xenopus oocytes. In addition, Harsh demonstrated uptake of glucosinolates tagged with a fluorescent label in yeast expressing GTR2. In parallel, Harsh took steps in establishing the SESAM technique for generating random mutant libraries of GTR2 to be screened in yeast for altered transport properties. In combination, this thesis lays a foundation for high-throughput screening of GTR2 mutant libraries, which will enable us to identify molecular determinants of transport properties
Harsh Yadav did his bachelor in Biotechnology at Shiv Nadar University, India before coming to Denmark for his master education at KU. He will continue as PhD student at Assoc. Prof. Hussam Nour-Eldin’s group. Hussam was also the main supervisor at the master project together with PhD student Christa Kanstrup.