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Team:Warsaw/Stage1/ExpressIt
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<h2>ExpressIt! - family of expression vectors with defined strength</h2> | <h2>ExpressIt! - family of expression vectors with defined strength</h2> | ||
| + | For your convenience we assembled a set of expression vectors. This year we prepared two tasty flavours: J23100 base and pT7 based vectors. Those promoters are available with 10 different RBS sequences (all that we could find in the registry). For constitutive protein expression use J32100 based vectors. T7promoter is stronger than J23100, but it is active only in strains harbouring T7 polymerase. Usually expression of T7polymerase is inducible therefore pT7 is a good choice for strong inducible promoter. For approximate guidelines about expression strength of our vectors see Figure 1. <br><br> | ||
| - | <h3>Key features:</h3> | + | |
| + | <h3>The Key features of ExpressIt - expression vectors kit: </h3> | ||
<ul> | <ul> | ||
<li><a href="https://2010.igem.org/Team:Warsaw/Stage1/RBSMeas">10 different measured RBS variants</a></li> | <li><a href="https://2010.igem.org/Team:Warsaw/Stage1/RBSMeas">10 different measured RBS variants</a></li> | ||
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<li>just one step to clone any protein coding BioBrick and you have expression construct ready</li> | <li>just one step to clone any protein coding BioBrick and you have expression construct ready</li> | ||
<li>wide spectrum of expression strengths</li> | <li>wide spectrum of expression strengths</li> | ||
| - | <li>ampicillin resistance | + | <li>ampicillin resistance - this is important for constructs that use T7 promoter. Why? |
| + | T7 promoter is active ONLY in presence of T7 polymerase. Therefore you have to use a <i>E.Coli</i> strain containing this gene. The most popular choice <i>E.coli</i> BL21, which has a chloramphenicol resistance gene in the genome.</li> | ||
<li>high copy ColE1 ori (based on <a href="http://partsregistry.org/Part:pSB1A2">pSB1A2</a> vector)</li> | <li>high copy ColE1 ori (based on <a href="http://partsregistry.org/Part:pSB1A2">pSB1A2</a> vector)</li> | ||
| - | <li>terminator included in vector sequence</li> | + | <li>terminator included in the vector sequence - <a href="http://partsregistry.org/Part:pSB1A2">pSB1A2</a> based.</li> |
</ul> | </ul> | ||
| - | + | <br> | |
<h3>Relative strength of ExpressIt! vector family:</h3> | <h3>Relative strength of ExpressIt! vector family:</h3> | ||
<img src="https://static.igem.org/mediawiki/2010/0/09/ExpressIt.png"> | <img src="https://static.igem.org/mediawiki/2010/0/09/ExpressIt.png"> | ||
| - | + | <br> | |
| + | Fig 1. The estimated expression vectors strength relative to pT7.B0034 - the strongest one. Estimation was based on The promoter and RBS measurement. Promoter and RBSes were assumed to be independent of each other (wchich is partly confirmed by our modeling results). | ||
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Latest revision as of 20:06, 26 October 2010
ExpressIt! - family of expression vectors with defined strength
For your convenience we assembled a set of expression vectors. This year we prepared two tasty flavours: J23100 base and pT7 based vectors. Those promoters are available with 10 different RBS sequences (all that we could find in the registry). For constitutive protein expression use J32100 based vectors. T7promoter is stronger than J23100, but it is active only in strains harbouring T7 polymerase. Usually expression of T7polymerase is inducible therefore pT7 is a good choice for strong inducible promoter. For approximate guidelines about expression strength of our vectors see Figure 1.The Key features of ExpressIt - expression vectors kit:
- 10 different measured RBS variants
- each variant in both constitutive (J23100 driven) and inducible (pT7 driven) version
- just one step to clone any protein coding BioBrick and you have expression construct ready
- wide spectrum of expression strengths
- ampicillin resistance - this is important for constructs that use T7 promoter. Why? T7 promoter is active ONLY in presence of T7 polymerase. Therefore you have to use a E.Coli strain containing this gene. The most popular choice E.coli BL21, which has a chloramphenicol resistance gene in the genome.
- high copy ColE1 ori (based on pSB1A2 vector)
- terminator included in the vector sequence - pSB1A2 based.
Relative strength of ExpressIt! vector family:
Fig 1. The estimated expression vectors strength relative to pT7.B0034 - the strongest one. Estimation was based on The promoter and RBS measurement. Promoter and RBSes were assumed to be independent of each other (wchich is partly confirmed by our modeling results).
