Difference between revisions of "Part:BBa K2084002"
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Regulatory ribosome binding site with T7 RNAP promoter.
 
Regulatory ribosome binding site with T7 RNAP promoter.
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<p>The switch is activated with a complementary RNA strand in the Collins paper. As shown in the figure below, it can also be activated with a complementary DNA strand, though protein expression is lower. Activation is measured by the expression of the reporter <i>lacZ</i> in absorbance at 570 nm.</p>
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    <img src="https://static.igem.org/mediawiki/2016/d/d2/T--Pittsburgh--Parts_PT7Toehold_DNAvRNA.jpg" style="display:block; margin:auto; width:50%">
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<p>The activation of the switch also increases with increasing concentration of trigger as shown below. Maximum activation is reached at a concentration of about 100 nM of DNA oligonucleotide trigger.</p>
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    <img src="https://static.igem.org/mediawiki/2016/f/f7/T--Pittsburgh--Parts_PT7Toehold_DNAtitration.jpg" style="display:block; margin:auto; width:50%">
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<p>In PURExpress, an <i>in vitro</i> protein synthesis reaction, maximum expression of the LacZ reporter protein regulated by the switch is reached by 2 hours.</p>
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    <img src="https://static.igem.org/mediawiki/2016/f/f1/T--Pittsburgh--Parts_PT7Toehold_Kinetics.jpg" style="display:block; margin:auto; width:60%">
  
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<h3>References</h3>
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<p>Pardee K, Green AA, Ferrante T, Cameron DE, DaleyKeyser A, Yin P, Collins JJ. <a href="http://www.cell.com/abstract/S0092-8674(14)01291-4" target="_blank">Paper-based synthetic gene networks</a>. <i>Cell</i> 159 (4): 940-954, 2014.</p>
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 17:38, 19 October 2016


pT7 Toehold

Regulatory ribosome binding site with T7 RNAP promoter.

The switch is activated with a complementary RNA strand in the Collins paper. As shown in the figure below, it can also be activated with a complementary DNA strand, though protein expression is lower. Activation is measured by the expression of the reporter lacZ in absorbance at 570 nm.

The activation of the switch also increases with increasing concentration of trigger as shown below. Maximum activation is reached at a concentration of about 100 nM of DNA oligonucleotide trigger.

In PURExpress, an in vitro protein synthesis reaction, maximum expression of the LacZ reporter protein regulated by the switch is reached by 2 hours.

References

Pardee K, Green AA, Ferrante T, Cameron DE, DaleyKeyser A, Yin P, Collins JJ. Paper-based synthetic gene networks. Cell 159 (4): 940-954, 2014.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]