Difference between revisions of "Part:BBa K2092002"

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<partinfo>BBa_K2092002 short</partinfo>
 
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The alcA promoter, P<i>alcA</i>, is originally found in <i>Aspergillus nidulans</i> as an ethanol regulon.  P<i>alcA</i> has a role in regulating the expression of gene <i>alcA</i>.  Gene <i>alcA</i> encodes for alcohol dehydrogenase I (ADHI) which facilitates the interconversion between alcohols and carbonyls.
  
The alcA promoter is one of the strongest inducible promoters in Aspergillus nidulans commonly used to overexpress proteins [1].  It has been shown that alcA promoter is also functional in monocotyledonous plant sugar cane [2] and Escherichia coli [3]. Its transciptional 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 AlcA promoter consists of 3 AlcR binding sites.  The number and position of the AlcR binding sites on the alcA promoter are crucial in determining its transcriptional activation strength.  It has also been shown that each AlcR target in the alcA promoter contributes differently to the activation of the downstream protein expression.
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===Usage and Biology===
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The 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 alcA promoter is also functional in monocotyledonous plant sugar cane [2] and <i>Escherichia coli</i> [3]. Its transciptional 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.
  
 
This part is an improved BioBrick previously characterised by 2011 iGEM DTU-Denmark. Sequencing results showed that two Prefix/Suffix restriction sites XbaI and SpeI were absent in the original part.  Hence the two missing restriction sites were added using PCR.  
 
This part is an improved BioBrick previously characterised by 2011 iGEM DTU-Denmark. Sequencing results showed that two Prefix/Suffix restriction sites XbaI and SpeI were absent in the original part.  Hence the two missing restriction sites were added using PCR.  
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[3] Hemmati, H. and Basu, C. (2015). Transcriptional analyses of an ethanol inducible promoter in Escherichia coli and tobacco for production of cellulase and green fluorescent protein. Biotechnology & Biotechnological Equipment, 29(6), 1043–1052.
 
[3] Hemmati, H. and Basu, C. (2015). Transcriptional analyses of an ethanol inducible promoter in Escherichia coli and tobacco for production of cellulase and green fluorescent protein. Biotechnology & Biotechnological Equipment, 29(6), 1043–1052.
 
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===Usage and Biology===
 
  
 
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Revision as of 15:42, 1 October 2016


PalcA, improved alcR inducible promoter from A. nidulans The alcA promoter, PalcA, is originally found in Aspergillus nidulans as an ethanol regulon. PalcA has a role in regulating the expression of gene alcA. Gene alcA encodes for alcohol dehydrogenase I (ADHI) which facilitates the interconversion between alcohols and carbonyls.


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]