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Team:Warsaw/Project
From 2011.igem.org
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<div class="note">Expression Adapters</div> | <div class="note">Expression Adapters</div> | ||
| - | <div> | + | <div align="justify"> |
So far all RBS parts was measured with GFP protein. We have measured the RBS parts with various fluorescent proteins and guess what - they are not as standard as we would like them to be. The strength of a RBS part depends on the protein used. Why - because the beginning of the protein influences the mRNA fold. When mRNA is strongly folded ribosome can not start translation. We came up with the idea of RBS parts fused with short 'protein beginnings' - we call this expression adapters. We developed an genetic algorithm to design expression adapters that would provide standardized protein expression from each RBS from community collection. Also we are working on adapters that increase expression of your favorite protein. Currently we are testing our design in the wet lab </div> | So far all RBS parts was measured with GFP protein. We have measured the RBS parts with various fluorescent proteins and guess what - they are not as standard as we would like them to be. The strength of a RBS part depends on the protein used. Why - because the beginning of the protein influences the mRNA fold. When mRNA is strongly folded ribosome can not start translation. We came up with the idea of RBS parts fused with short 'protein beginnings' - we call this expression adapters. We developed an genetic algorithm to design expression adapters that would provide standardized protein expression from each RBS from community collection. Also we are working on adapters that increase expression of your favorite protein. Currently we are testing our design in the wet lab </div> | ||
<br> | <br> | ||
<div class="note">Synthetic Cloning</div> | <div class="note">Synthetic Cloning</div> | ||
| - | <div>Our goal is to make easy and efficient protocol for cell free | + | <div align="justify">Our goal is to make easy and efficient protocol for cell free |
cloning. It skips most basic step of classical cloning approach - | cloning. It skips most basic step of classical cloning approach - | ||
plasmid propagation in bacteria or yeast. This allows cloning of | plasmid propagation in bacteria or yeast. This allows cloning of | ||
Revision as of 12:09, 18 September 2011
Synthetic cloning and expression control
This year's goal of iGEM Warsaw team is to develop 2 foundational
techniques for synthetic biologists:
Expression Adapters
So far all RBS parts was measured with GFP protein. We have measured the RBS parts with various fluorescent proteins and guess what - they are not as standard as we would like them to be. The strength of a RBS part depends on the protein used. Why - because the beginning of the protein influences the mRNA fold. When mRNA is strongly folded ribosome can not start translation. We came up with the idea of RBS parts fused with short 'protein beginnings' - we call this expression adapters. We developed an genetic algorithm to design expression adapters that would provide standardized protein expression from each RBS from community collection. Also we are working on adapters that increase expression of your favorite protein. Currently we are testing our design in the wet lab
Synthetic Cloning
Our goal is to make easy and efficient protocol for cell free
cloning. It skips most basic step of classical cloning approach -
plasmid propagation in bacteria or yeast. This allows cloning of
constructs that are toxic to host cells (i.e. nucleases or lysins) and
speeds up the whole procedure at least three times. The cell free
cloning process involves rolling circle amplification of ligation
products by phi29 polymerase. Our protocol is speed-oriented so we'll
try to optimize all time consuming steps: DNA digestion, substrate
purification, ligation and product amplification. We'll also try to
deal with specificity problems of phiX29 polymerase by designing RNA
primers optimized for BioBrick amplification.
