A major part of our work involved direct use of our target protein molecule, the Tau protein. We had planned to produce the protein ourselves. With protein production out of the way, the big challenge that remained was its phosphorylation, the post-translational modification of the Tau protein.
To resolve this issue, we consulted a molecular biologist, Prof. Tanweer Hussain, Developmental Biology and Genetics, IISc. Prof. Hussain, who is closely associated with Nobel Laureate Prof. Venkatraman Ramakrishnan, is a pioneer in the field of RNA biology. Upon consultation, we came to learn about the difficulty and the necessity of experience for mammalian expression of protein.
After a series of consecutive discussion with Prof. Varadarajan and Prof. Hussain, we decided to use a bacterial model for protein expression and phosphorylate the purified protein in-vitro.