Difference between revisions of "Part:BBa K3350858"
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Several transcriptional regulatory protein families are involved in the degradation of aromatic compounds in bacteria, differing both in their structure and in the activation or repression mechanisms. LysR-type transcriptional regulators (LTTRs), are characterized by an N-terminal DNA-binding helix-turn-helix motif and a C-terminal co-inducer-binding domain. According to Tropel and V an Der Meer, all LTTRs involved in aromatic compounds degradation pathways act as transcriptional activators with the co-inducer being either the degraded compound or an intermediate in the degradation pathway. YhaJ protein, a member of the LysR type family, acts as a transcriptional regulator of yqjF activation (BBa_K3350857). We perform random error-prone PCR on the sequence to improve the response of yqjF promoter to DNT.
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Several transcriptional regulatory protein families are involved in the metabolism of aromatic compounds in bacteria, and have difference in both their structures and in the activation mechanisms. The LysR-type transcriptional regulators (LTTRs) are characterized by an N-terminal DNA-binding helix-turn-helix motif and a C-terminal co-inducer-binding domain. According to Tropel and V an Der Meer, all LTTRs involved in aromatic compounds degradation pathways can act as transcriptional activators with the co-inducers, which are either degraded compounds or intermediates in the degradation pathways. The YhaJ protein, a member of the LysR type family, acts as a transcriptional regulator to activate the the yqjF promoter (BBa_K1316002). We had optimized the yqjF promoter to obtain its new version yqjF3rd (BBa_K3350862). Thus, when we overexpressed the yhaJ gene together with an yqjF3rd-containing reporter construct in the engineered bacteria, we could reduce the DNT detection threshold of the yqjF promoter from 25 mg/L to 0.25 mg/L, which is a 100-fold increase of the sensitivity.
  
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 12:49, 24 October 2020


yhaJ1st (DNA-binding transcriptional activator)
Several transcriptional regulatory protein families are involved in the metabolism of aromatic compounds in bacteria, and have difference in both their structures and in the activation mechanisms. The LysR-type transcriptional regulators (LTTRs) are characterized by an N-terminal DNA-binding helix-turn-helix motif and a C-terminal co-inducer-binding domain. According to Tropel and V an Der Meer, all LTTRs involved in aromatic compounds degradation pathways can act as transcriptional activators with the co-inducers, which are either degraded compounds or intermediates in the degradation pathways. The YhaJ protein, a member of the LysR type family, acts as a transcriptional regulator to activate the the yqjF promoter (BBa_K1316002). We had optimized the yqjF promoter to obtain its new version yqjF3rd (BBa_K3350862). Thus, when we overexpressed the yhaJ gene together with an yqjF3rd-containing reporter construct in the engineered bacteria, we could reduce the DNT detection threshold of the yqjF promoter from 25 mg/L to 0.25 mg/L, which is a 100-fold increase of the sensitivity.

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]