Evaluation of Myristoylation for Post-Translational Modification in a Coupled Transcription/Translation Protein Synthesis Environment

In vivo myristoylation has been used to post-translationally modify proteins. The apparent lack of specificity of the N-myristoyltransferace for myristate has been used to attach myristoyl-like entities with unique functionality. For example a myristoyl-like molecule containing a terminal azide has been attached to proteins by N-myristoyltransferase. We seek evaluate the ability to use myristoylation to post-translationally modify proteins expressed in a Cell-free protein synthesis environment. Specifically, we seek to optimize the expression and concentrations of the N-myristoyltransferase, myristate, and the modified proteins to compare the efficiency of a cell-free approach to in vivo myristoylation.