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National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
Biophysical characterization of protein libraries composed of different amino acid repertoires
 Neuwirthová, Tereza ; Hlouchová, Klára (advisor) ; Ptáček, Jakub (referee)
This study is part of a project which aims to understand evolution of genetic code together with structural and functional analysis of prebiotic proteins. The repertoire of amino acids in the first proteins was probably developing in time and it influenced the development of structure and function of today's proteins. First amino acid alphabet was apparently only half of the size of present alphabet, which contains twenty amino acids. These ten amino acids were probably prebiotically available from endogenous and exogenous sources. This work includes cell-free expression and purification of two randomized protein libraries (containing approximately 1011 variants) with various amino acid composition and following comparison of their propensity to form secondary (using circular dichroism) and tertiary (using proteolytical analysis of sequences) structures. First library contains only ten probably prebiotically available amino acids; second library contains all twenty amino acids in today's genetic code. This project could help us understand benefits of genetic code expansion in terms of developing structure in protein sequences. The whole research could theoretically contribute a few basic questions not only in the fields of protein evolution but also in areas of synthetic biology or protein...
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Cell-free expression of protein libraries made of different amino acid repertoires
 Neuwirthová, Tereza ; Hlouchová, Klára (advisor) ; Míšek, Jiří (referee)
This bachelor thesis is an introduction to a project dealing with the influence of genetic code evolution (and the consequentamino acid repertoire in proteins) on the evolution of protein structure and function. The initial amino acid alphabet was apparently only half the size of the present alphabet and according to available metaanalyses it included only about 10 prebiotically present amino acids derived from potential exogenous or endogenous sources. This work includes mRNA transcription and cell-free expression of five randomized protein libraries (containing approximately 1011 variants). These libraries differ in their amino acid composition and reflect individual predicted stages of genetic code evolution. The main goal is to convert libraries from DNA to protein form to provide a source for library purification and their subsequent biophysical (occurrence of secondary and tertiary structure) and functional analyses. The whole research could theoretically contribute to a few basic questions not only in the fields of protein evolution but also in areas of synthetic biology or protein engineering. Keywords: Cell-free expression, proteins, genetic code evolution This thesis is written in Czech.
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