Difference between revisions of "Team:CityU HK/Description"

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<h2 class="wsite-content-title" style="text-align:left;">How are we going to tackle it?</h2>
 
<h2 class="wsite-content-title" style="text-align:left;">How are we going to tackle it?</h2>
  
<div class="paragraph" style="text-align:left;"><span style="">We aim to design and construct a probiotic&nbsp;<em style="">E. coli</em>&nbsp;strain that (1) produces high level of the beta-galactosidase enzyme (a type of lactase) constitutively, and (2) autolyses to release the beta-galactosidase enzyme upon detecting lactose in the external environment. For process (1), we will construct a&nbsp;<em style="">lacZ</em>&nbsp;(beta-galactosidase) &nbsp;-<em style="">lacY</em>&nbsp;(lactose permease) biobrick that is driven by a strong constitutive promoter &nbsp;to enable high &nbsp;expression of the beta-galactosidase enzyme. For process (2), we will construct a lysis cassette&nbsp; consisting of the S (holin)-R (endolysin) -Rz (spannin) genes driven by a&nbsp;<em style="">lac</em>I promoter&nbsp; to facilitate lactose-induced cell lysis.</span><br /><br /><span style="">To optimize the efficiency of&nbsp;<em style="">E. coli</em>&nbsp;cell lysis, we will carry out the following modifications to certain genes in the lysis cassette:</span><br /><span style=""></span><span style="">&nbsp; &nbsp; &nbsp;1. Genes in the lysis cassette &nbsp;will be codon optimized for optimal expression in&nbsp;<em style="">E. coli;</em></span><br /><span style=""></span><span style="">&nbsp; &nbsp; &nbsp;2. The anti-holin component will be deleted to speed up cell lysis;</span><br /><span style=""></span><span style="">&nbsp; &nbsp; &nbsp;3. Specific mutations will be introduced into the lysis gene cassette to facilitate shorter lysis time.</span><br /><span style=""></span><br /><span style="">In this project, we will also prepare and compare the lysis efficiency of two different lysis cassettes &ndash; (1) Cassette 1 will be constructed using the S</span>&lambda;<span style="">, R</span>&lambda;<span style="">&nbsp;and Rz</span>&lambda;<span style="">&nbsp;genes derived from&nbsp;</span><em style="">E. coli&nbsp;</em><span style="">lambda phage, and (2) Cassette 2 will be constructed using the S</span>21<span style="">, R</span>21<span style="">&nbsp;and Rz</span>21<span style="">&nbsp;genes derived from&nbsp;</span><em style="">E. coli&nbsp;</em><span style="">phage 21.</span></div></div>
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<div class="paragraph" style="text-align:left;"><span style="">We aim to design and construct a probiotic&nbsp;<em style="">E. coli</em>&nbsp;strain that (1) produces high level of the beta-galactosidase enzyme (a type of lactase) constitutively, and (2) autolyses to release the beta-galactosidase enzyme upon detecting lactose in the external environment. For process (1), we will construct a&nbsp;<em style=""><i>lacZ</i></em>&nbsp;(beta-galactosidase) &nbsp;-<em style=""><i>lacY'</i></em>&nbsp;(lactose permease) biobrick that is driven by a strong constitutive promoter &nbsp;to enable high &nbsp;expression of the beta-galactosidase enzyme. For process (2), we will construct a lysis cassette&nbsp; consisting of the S (holin)-R (endolysin) -Rz (spannin) genes driven by a&nbsp;<em style="">lac</em>I promoter&nbsp; to facilitate lactose-induced cell lysis.</span><br /><br /><span style="">To optimize the efficiency of&nbsp;<em style="">E. coli</em>&nbsp;cell lysis, we will carry out the following modifications to certain genes in the lysis cassette:</span><br /><span style=""></span><span style="">&nbsp; &nbsp; &nbsp;1. Genes in the lysis cassette &nbsp;will be codon optimized for optimal expression in&nbsp;<em style="">E. coli;</em></span><br /><span style=""></span><span style="">&nbsp; &nbsp; &nbsp;2. The anti-holin component will be deleted to speed up cell lysis;</span><br /><span style=""></span><span style="">&nbsp; &nbsp; &nbsp;3. Specific mutations will be introduced into the lysis gene cassette to facilitate shorter lysis time.</span><br /><span style=""></span><br /><span style="">In this project, we will also prepare and compare the lysis efficiency of two different lysis cassettes &ndash; (1) Cassette 1 will be constructed using the S</span>&lambda;<span style="">, R</span>&lambda;<span style="">&nbsp;and Rz</span>&lambda;<span style="">&nbsp;genes derived from&nbsp;</span><em style="">E. coli&nbsp;</em><span style="">lambda phage, and (2) Cassette 2 will be constructed using the S</span>21<span style="">, R</span>21<span style="">&nbsp;and Rz</span>21<span style="">&nbsp;genes derived from&nbsp;</span><em style="">E. coli&nbsp;</em><span style="">phage 21.</span></div></div>
 
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Revision as of 12:18, 13 September 2015

Description - iGEM2015 wiki