Difference between revisions of "Part:BBa K2092003"

 
Line 1: Line 1:
 +
__NOTOC__
 +
<partinfo>BBa_K2092002 short</partinfo>
  
 +
P<i>alcA</i> is one of the strongest inducible promoters in <i>Aspergillus nidulans</i> commonly used to overexpress proteins [1].  It has been shown that P<i>alcA</i> promoter is also functional in monocotyledonous plant sugar cane [2] and <i>Escherichia coli (E.coli)</i>[3]. Its transcriptional activation is dependent on the binding of its positive transcriptional regulator AlcR with various substrates that employ a hydroxyl group, for example ethanol and threonine.  The native P<i>alcA</i> consists of 3 AlcR binding sites.  The number and position of the AlcR binding sites on the P<i>alcA</i> are crucial in determining its transcriptional activation strength.  It has also been shown that each AlcR target in the P<i>alcA</i> contributes differently to the activation of the downstream protein expression [1].
 +
 +
 +
This part is a variant of P<i>alcA</i> <a href=https://parts.igem.org/Part:BBa_K2092002>(BBa_K2092002)</a>.  In contrast to the native construct (three AlcR binding sites <i>abc</i>), this variant consists only two binding sites <i>bc</i>. Previous study on sugarcane has shown that there is a 70% decrease in promoter strength for P<i>alcA</i> with only binding sites <i>bc</i> [2].  This finding, however, has not been verified using <i>E.coli</i> as a chassis.  This then forms the very basis of our project. 
 +
 +
 +
 +
 +
 +
<!-- -->
 +
<span class='h3bb'>Sequence and Features</span>
 +
<partinfo>BBa_K2092002 SequenceAndFeatures</partinfo>
 +
 +
 +
<!-- Uncomment this to enable Functional Parameter display
 +
===Functional Parameters===
 +
<partinfo>BBa_K2092002 parameters</partinfo>
 +
<!-- -->

Revision as of 16:41, 1 October 2016

PalcA, improved alcR inducible promoter from A. nidulans

PalcA is one of the strongest inducible promoters in Aspergillus nidulans commonly used to overexpress proteins [1]. It has been shown that PalcA promoter is also functional in monocotyledonous plant sugar cane [2] and Escherichia coli (E.coli)[3]. Its transcriptional activation is dependent on the binding of its positive transcriptional regulator AlcR with various substrates that employ a hydroxyl group, for example ethanol and threonine. The native PalcA consists of 3 AlcR binding sites. The number and position of the AlcR binding sites on the PalcA are crucial in determining its transcriptional activation strength. It has also been shown that each AlcR target in the PalcA contributes differently to the activation of the downstream protein expression [1].


This part is a variant of PalcA <a href=https://parts.igem.org/Part:BBa_K2092002>(BBa_K2092002)</a>. In contrast to the native construct (three AlcR binding sites abc), this variant consists only two binding sites bc. Previous study on sugarcane has shown that there is a 70% decrease in promoter strength for PalcA with only binding sites bc [2]. This finding, however, has not been verified using E.coli as a chassis. This then forms the very basis of our project.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 274
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]