<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li>

<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li>

 

<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a>

                                </ul></li>  

There are many challenges for anyone who wants to do synthetic biology in a new less common organism, like getting familiar with new protocols and techniques. But another major problem is that there is a lack of fundamental parts available like plasmids, reporter genes and promoters. A big part of our project has been to develop these basic parts and make them biobrick compatible. This is done in order to open up the Lactobacillus genus as the new probiotic chassi of choice for the iGEM community. We also intend to use it ourselves for doing metabolic engineering in Lactobacillus. For this we have worked to create two <a href="https://2013.igem.org/Team:Uppsala/vectors"> shuttle vectors </a>, a library of <a href="https://2013.igem.org/Team:Uppsala/promoters"> constitutive promoters </a> and a couple of <a href="https://2013.igem.org/Team:Uppsala/reporter-genes"> reporter genes </a> that should work in both E. coli and Lactobacillus. Because E. coli is substantially easier and faster to work with the intention is to allow construction and some characterisation to be done in E. coli and then in the end transfer the complete construct to lactobacillus.  

<div id="promoter_text"> Different constructs need different levels of expression. We have constructed and characterised a library of constitutive promoters of different strengths. These are based on a consensus sequence from promoters in Lactococcus lactis but have been shown to also work very well in E. coli and Lactobacillus.  

<div id="reporter_text"> Many techniques and methods require fluorescent reporter genes to function. While some fluorescent proteins works poorly in Lactobacillus we have biobricked a codon-optimized version of mCherry that has been shown to work. This has been done according to assembly standard 25 to allow it to be used as a fusion protein.  

<div id="vector_text"> Perhaps the most important chassi parts we have constructed are shuttle vectors that work both in Lactobacillus and E. coli. Because Lactobacillus is significantly harder to work with and ligations are hard to transform, constructs should first be assembled and prepped in E. coli and then transferred to Lactobacillus with the shuttle vector.  

 

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