Cell-Free Protein Systems from Yersinia pestis are Functional and Growth-Temperature Dependent

Cellular lysates capable of transcription and translation have become valuable tools for prototyping genetic circuits, screening engineered functional parts, and producing biological components. Here we report that lysates derived from Yersinia pestis CO92(-) are functional and can utilize both the E. coli sigma 70 and the bacteriophage T7 promoter systems to produce green fluorescent protein (GFP). Because of the natural lifestyle of Y. pestis, lysates were produced from cultures grown at 21 degrees C, 26 degrees C, and 37 degrees C to mimic the infection cycle. Regardless of the promoter system the GFP production from 37 degrees C was the most productive and the 26 degrees C lysate was the least. When reactions are initiated with 5 nM of DNA, the GFP output of the 37 degrees C lysate is comparable with the productivity of other non-E. coli systems. The data we present demonstrate that, without genetic modification to enhance productivity, cell-free extracts from Y. pestis are functional and dependent on the temperature at which the bacterium was grown.