Difference between revisions of "Part:BBa K3198002"

 
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<partinfo>BBa_K3198002 short</partinfo>
 
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<!-- This part contains the toxin component of a type II toxin-antitoxin (TA) system. HicA is a probable translation-independent mRNA interferase.<br><br> -->
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3198002 SequenceAndFeatures</partinfo>
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===Usage===
 
It is demonstrated that researchers that the RES-Xre locus from Photorhabdus luminescens and other bacterial species function as bona fide TA modules in Escherichia coli.  
 
It is demonstrated that researchers that the RES-Xre locus from Photorhabdus luminescens and other bacterial species function as bona fide TA modules in Escherichia coli.  
 
The activation of the toxin in vivo causes a depletion of intracellular NAD+ levels, eventually leading to inhibition of cell growth in E. coli and inhibition of global macromolecular biosynthesis.  
 
The activation of the toxin in vivo causes a depletion of intracellular NAD+ levels, eventually leading to inhibition of cell growth in E. coli and inhibition of global macromolecular biosynthesis.  
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===Biology===
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===Usage and Biology===
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This part is from the res-xre locus from Photorhabdus luminescens and other bacterial species.
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===Characterisation===
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<!-- Team NUS 2019 has added a new biobrick (BBa_K3198000) into the iGEM repository this year. This biobrick was found to possess a bacteriostatic effect as reported by Gerdes et al in 2008 (doi:10.1128/JB.01013-08) and was therefore used by Team NUS 2019 as part of their sleep-wake module to control the growth of E. coli in their project.
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<br><br>(BBa_K3198000) was placed under an IPTG-inducible promoter and various IPTG concentrations were utilized to determine their effect on the growth of native <i>MG1655</i>. In the same plasmid, another cassette containing GFP reporter gene under constitutive promoter was present to enable characterization of the effect of HicA on protein production. 
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<br><br>Characterization of cells transformed with this plasmid was performed at 37°C for 12h continuously. The results showed that IPTG concentrations of 100μM, 500μM and 2mM resulted in growth arrest as shown by a reduction and plateau in OD600. IPTG concentrations beyond 500μM did not show further reduction in OD600.
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<br><br>Figure 1: Growth curve of control <i>MG1655</i> unaffected by IPTG
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<br><br>Figure 2: Growth curve of <i>MG1655</i> transformed with HicA-containing plasmid
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<br><br>Furthermore, team NUS 2019 also studied the effect of (BBa_K3198000) on protein expression. The results demonstrated that HicA-containing cells when induced with IPTG resulted in a drop in total GFP level, as opposed to uninduced HicA-containing cells. This suggests that the effect of (BBa_K3198000) on cell growth is likely to affect its protein expression ability.
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<br><br>Figure 3: Total GFP curve of HicA-plasmid containing cells with different IPTG induction (0M, 100μM, 500μM and 2mM)
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<br><br>In summary, we believe that (BBa_K3198000) is a new BioBrick capable of causing cell growth arrest and suppressing protein expression. -->
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===References===
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Skjerning, R. B., Senissar, M., Winther, K. S., Gerdes, K., & Brodersen, D. E. (2018). The RES domain toxins of RES-Xre toxin-antitoxin modules induce cell stasis by degrading NAD . Molecular Microbiology, 111(1), 221–236. doi: 10.1111/mmi.14150
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Milunovic, B., diCenzo, G.C., Morton, R.A.and Finan, T.M. (2014) Cell growth inhibition upon deletion of four toxin‐antitoxin loci from the megaplasmids of Sinorhizobium meliloti. Journal of Bacteriology, 196, 811–824.
  
 
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<span class='h3bb'>Sequence and Features</span>
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<!-- Uncomment this to enable Functional Parameter display  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3198002 parameters</partinfo>
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<partinfo>BBa_K3198001 parameters</partinfo>
 
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Revision as of 09:51, 31 August 2019


RES



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]

Usage

It is demonstrated that researchers that the RES-Xre locus from Photorhabdus luminescens and other bacterial species function as bona fide TA modules in Escherichia coli. The activation of the toxin in vivo causes a depletion of intracellular NAD+ levels, eventually leading to inhibition of cell growth in E. coli and inhibition of global macromolecular biosynthesis.


Biology

This part is from the res-xre locus from Photorhabdus luminescens and other bacterial species.


Characterisation

References

Skjerning, R. B., Senissar, M., Winther, K. S., Gerdes, K., & Brodersen, D. E. (2018). The RES domain toxins of RES-Xre toxin-antitoxin modules induce cell stasis by degrading NAD . Molecular Microbiology, 111(1), 221–236. doi: 10.1111/mmi.14150

Milunovic, B., diCenzo, G.C., Morton, R.A.and Finan, T.M. (2014) Cell growth inhibition upon deletion of four toxin‐antitoxin loci from the megaplasmids of Sinorhizobium meliloti. Journal of Bacteriology, 196, 811–824.