Investigation of Protein Dynamics Using Genetically Encoded Unnatural Amino Acids

Srivastava, A K (2015) Investigation of Protein Dynamics Using Genetically Encoded Unnatural Amino Acids. Master's Thesis / Essay, Biology.

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Abstract

Protein engineering has become an extensively used tool in many fields which allow us to study protein functions and characterize proteins using range of available biophysical methods. With the genetic code expansion, it is now possible to incorporate functional groups and other properties in the proteins which are naturally not present. Site specific incorporation of unnatural amino acid (uAA) is widely used in protein engineering. However, incorporation of multiple uAAs remains a significant challenge. Tunability of protein expression for multiple uAAs incorporation using orthogonal ribosome remains another important challenge. In this study we try to investigate the role of inducible promoter in tunability of protein expression using orthogonal ribosome and orthogonal ribosome binding sites along with tRNA/amino acyl RNA synthetase pair. Six different o-RBS were designed based on the work of Rackham and Chin [26] during the study on an inducible promoter. 4 best fit o-RBS were selected after screening experiment. Expression of protein incorporating uAA was done with orthogonal ribosome. Unfortunately, the experiment failed. Further analysis on failure concluded that the functionality of orthogonal ribosome was lost due to some unexplained reason.

Item Type:	Thesis (Master's Thesis / Essay)
Degree programme:	Biology
Thesis type:	Master's Thesis / Essay
Language:	English
Date Deposited:	15 Feb 2018 08:06
Last Modified:	15 Feb 2018 08:06
URI:	https://fse.studenttheses.ub.rug.nl/id/eprint/13119

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