Difference between revisions of "Part:BBa K3970011"
 
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The BS module，which is the lexO is the diversification and characterization of the LexA-binding motifs. We find it in an paper.
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__NOTOC__
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The previously characterized LexA binding motif has a consensus sequence of a 16 bp palindrome, CTGTATATATATACAG. Yeast contains a number of genes regulated by the LexA repressor. These genes, as well as the previously used modified GAL1 promoter with 8 LexA BS, contain a spectrum of sequence modifications deviating from the consensus LexA binding site .Based on the native LexA binding sites present in the regulatory region of the lexA gene and in the ColEI operator, the author designed versions of the LexA BS with minor sequence modifications and tested their binding affinity to purified sTF16 in the electrophoretic mobility shift assay (EMSA) . The sTF16 used was composed of a LexA DNA binding domain, the Herpes simplex virus transactivation domain (VP16) and a 6×His-tag. The results indicate that all four versions of the BSs bind similarly to the LexA-DNA-binding domain in vitro and therefore can also be expected to perform similarly within a synthetic output promoter in vivo.
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<partinfo>BBa_K3970011 short</partinfo>
  
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Reference:
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==Usage and Biology==
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The BS module，which is the lexO is the diversification and characterization of the LexA-binding motifs. We find it in an paper.<br>
 +
The previously characterized LexA binding motif has a consensus sequence of a 16 bp palin
 +
drome, CTGTATATATATACAG. Yeast contains a number of genes regulated by the
 +
LexA repressor. These genes, as well as the previously used modified GAL1 promoter with 8
 +
LexA BS, contain a spectrum of sequence modifications deviating from the consensus LexA
 +
binding site .Based on the native LexA binding sites present in the regulatory region of
 +
the lexA gene and in the ColEI operator, the author designed versions of the LexA BS with
 +
minor sequence modifications and tested their binding affinity to purified sTF16 in the electrophoretic mobility shift assay (EMSA) . The sTF16 used was composed of a LexA DNA binding domain, the Herpes simplex virus transactivation domain (VP16) and a 6×His-tag. The results indicate that all four versions of the BSs bind similarly to the LexA-DNA-binding domain in vitro and therefore can also be expected to perform similarly within a synthetic output promoter in vivo.
 +
 
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==Reference==
 
Synthetic Transcription Amplifier System for Orthogonal Control of Gene Expression in Saccharomyces cerevisiae;PLOS ONE | DOI:10.1371/journal.pone.0148320
 
Synthetic Transcription Amplifier System for Orthogonal Control of Gene Expression in Saccharomyces cerevisiae;PLOS ONE | DOI:10.1371/journal.pone.0148320
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3970009 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K3970009 parameters</partinfo>
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Latest revision as of 12:10, 8 October 2021

LexO

Usage and Biology

The BS module，which is the lexO is the diversification and characterization of the LexA-binding motifs. We find it in an paper.
The previously characterized LexA binding motif has a consensus sequence of a 16 bp palin drome, CTGTATATATATACAG. Yeast contains a number of genes regulated by the LexA repressor. These genes, as well as the previously used modified GAL1 promoter with 8 LexA BS, contain a spectrum of sequence modifications deviating from the consensus LexA binding site .Based on the native LexA binding sites present in the regulatory region of the lexA gene and in the ColEI operator, the author designed versions of the LexA BS with minor sequence modifications and tested their binding affinity to purified sTF16 in the electrophoretic mobility shift assay (EMSA) . The sTF16 used was composed of a LexA DNA binding domain, the Herpes simplex virus transactivation domain (VP16) and a 6×His-tag. The results indicate that all four versions of the BSs bind similarly to the LexA-DNA-binding domain in vitro and therefore can also be expected to perform similarly within a synthetic output promoter in vivo.

Reference

Synthetic Transcription Amplifier System for Orthogonal Control of Gene Expression in Saccharomyces cerevisiae;PLOS ONE | DOI:10.1371/journal.pone.0148320


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]