Team:Austin Texas/kit

From 2014.igem.org

(Difference between revisions)
Line 86: Line 86:
<h2>Motivation</h2>
<h2>Motivation</h2>
-
[[File:Translation of ncAA 10-16-14.png|300px|thumb|right|<b>Figure 1</b> Translation of non-canonical amino acids.]]
+
[[File:Translation of ncAA 10-16-14.png|300px|thumb|right|<b>Figure 1.</b> The process of charging a ncAA, followed by its incorporation during translation.  To do this successfully, a novel tRNA synthetase must be evolved that specifically recognizes the ncAA.]]
In order to recode UAG, a synthetase must be mutated to effectively "charge" a ncAA onto the corresponding tRNA. Various methods of directed evolution are typically used to modify a synthetase such that it can interact with and then charge a specific ncAA (Liu et al. 2010). The ncAA synthetases available have ranging levels of reported efficiency and are not well characterized.  Many of the ncAA are not widely used, they are published in short articles lacking full documentation, and our own unpublished experiences of working with a number of ncAAs indicate that not all ncAA synthetases are created equal. Thus, we created a standard kit designed to characterize the properties of any ncAA synthetase/tRNA pair.  Our goal is to produce a standardized kit that is cheap, reproducible, easy to use, AND easily portable (i.e. you don't need a lot of advanced equipment).
In order to recode UAG, a synthetase must be mutated to effectively "charge" a ncAA onto the corresponding tRNA. Various methods of directed evolution are typically used to modify a synthetase such that it can interact with and then charge a specific ncAA (Liu et al. 2010). The ncAA synthetases available have ranging levels of reported efficiency and are not well characterized.  Many of the ncAA are not widely used, they are published in short articles lacking full documentation, and our own unpublished experiences of working with a number of ncAAs indicate that not all ncAA synthetases are created equal. Thus, we created a standard kit designed to characterize the properties of any ncAA synthetase/tRNA pair.  Our goal is to produce a standardized kit that is cheap, reproducible, easy to use, AND easily portable (i.e. you don't need a lot of advanced equipment).

Revision as of 16:28, 17 October 2014