Team:Purdue/Project/BCD

From 2013.igem.org

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<p>Despite having progressed extensively in the field of synthetic biology in terms of DNA synthesis, analysis and transplanting, we still cannot reliably, quantitatively measure expression of new genetic constructs. We engineered an expression cassette to control transcription and translation initiation which can be reused in new genetic contexts. The Bicistronic design(BCD) consists of two Shine-Dalgarno sequences in its translation element which when combined with indiscriminate gene of interests are known to reliably express within twofold of the relative target expression window.</p>
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<p>Within the last few decades, DNA synthesis, sequencing and recombination technologies have significantly increased in sophistication and in turn practicality. However, the unpredictable variances in the behavior of standardized parts across setting changes remain an obstacle to time-efficient, rational gene network design. Vivek Mutalik and colleagues have in the past year designed bicistronic expression operating units (EOUs). Using these bicistronic designs (BCDs) Mutalik and others managed to express arbitrary genes within a twofold target window with ~93% reliability (Mutalik, 2012). The next best system at the time could only do the same with ~47% reliability (Salis, 2009). This summer, we attempted to make four existing BCD designs BioBrick compatible and modify them for assembly in an all-in-one-pot Golden Gate Cloning Reaction using the type IIs restriction enzyme BpiI.</p>
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Revision as of 15:29, 27 September 2013


PurdueLogo2013.png

Bicistronic Design

Improving and Regulating Protein Expression

Overview

Within the last few decades, DNA synthesis, sequencing and recombination technologies have significantly increased in sophistication and in turn practicality. However, the unpredictable variances in the behavior of standardized parts across setting changes remain an obstacle to time-efficient, rational gene network design. Vivek Mutalik and colleagues have in the past year designed bicistronic expression operating units (EOUs). Using these bicistronic designs (BCDs) Mutalik and others managed to express arbitrary genes within a twofold target window with ~93% reliability (Mutalik, 2012). The next best system at the time could only do the same with ~47% reliability (Salis, 2009). This summer, we attempted to make four existing BCD designs BioBrick compatible and modify them for assembly in an all-in-one-pot Golden Gate Cloning Reaction using the type IIs restriction enzyme BpiI.

Background

Experimental Design

Data

Results and Conclusion

Future Work