Difference between revisions of "Team:IISc-Bangalore/Design"

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<h1 id="t7-expression-system">T7 expression system</h1>
 
<h1 id="t7-expression-system">T7 expression system</h1>
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<partinfo>BBa_K2319000 SequenceAndFeatures</partinfo>
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<p>Our third method to induce gas vesicle aggregation (SpyCatcher-SpyTag binding) involves protein overexpression, and no system is better than <i>E. coli</i> strain BL21 (DE3)'s T7 expression system for this purpose: BL21 (DE3) is deficient in <i>lon</i> and <i>ompT</i> proteases. BL21 (DE3) has the T7 RNA polymerase gene integrated into its genome under the lac operon; adding IPTG induces expression of T7 RNA polymerase, which recognizes the T7 promoter sequence. Any gene inserted downstream of the T7 promoter can thus be expressed.</p>
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<p>Using <a href="http://parts.igem.org/Part:BBa_K525998">BBa_K525998</a> (T7 promoter+RBS) and  </p>
 
<p>Using <a href="http://parts.igem.org/Part:BBa_K525998">BBa_K525998</a> (T7 promoter+RBS) and  </p>
  

Revision as of 12:09, 30 October 2017

  1. T7 expression system
  2. sfGFP-SpyCatcher
  3. mCherry-SpyTag
  4. GvpC fusion

T7 expression system

Our third method to induce gas vesicle aggregation (SpyCatcher-SpyTag binding) involves protein overexpression, and no system is better than E. coli strain BL21 (DE3)'s T7 expression system for this purpose: BL21 (DE3) is deficient in lon and ompT proteases. BL21 (DE3) has the T7 RNA polymerase gene integrated into its genome under the lac operon; adding IPTG induces expression of T7 RNA polymerase, which recognizes the T7 promoter sequence. Any gene inserted downstream of the T7 promoter can thus be expressed.

Using BBa_K525998 (T7 promoter+RBS) and

sfGFP-SpyCatcher

mCherry-SpyTag

GvpC fusion