Difference between revisions of "Part:BBa K3198003"

 
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<partinfo>BBa_K3198003 short</partinfo>
 
<partinfo>BBa_K3198003 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_K3198003 SequenceAndFeatures</partinfo>
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===Usage===
 
It is demonstrated 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 the res-xre locus from Photorhabdus luminescens and other bacterial species function as bona fide TA modules in Escherichia coli.  
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<br><br>
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It was shown that a RES toxin encoded by the α‐proteobacterium, <i>Sinorhizobium meliloti</i>, is toxic when expressed in Escherichia coli and can be counteracted by its cognate Xre antitoxin.
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<br><br>
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RES toxin from several bacterial species reversibly inhibits cell growth in <i>E. coli</i>, and that expression of the Xre antitoxin neutralizes this toxicity. However, the cellular function of the RES toxin when activated, as well as structure and the antitoxic mechanism of the Xre antitoxin, remain unknown.
  
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it was shown that a RES toxin encoded by the &#945;&#8208;proteobacterium, Sinorhizobium meliloti, is toxic when expressed in Escherichia coli and can be counteracted by its cognate Xre antitoxin.
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===Biology===
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RES toxin from several bacterial species reversibly inhibits cell growth in E. coli, and that expression of the Xre antitoxin neutralizes this toxicity. However, the cellular function of the RES toxin when activated, as well as structure and the antitoxic mechanism of the Xre antitoxin, remain unknown.
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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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===Usage and Biology===
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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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<br><br>
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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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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3198003 SequenceAndFeatures</partinfo>
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3198003 parameters</partinfo>
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<partinfo>BBa_K3198001 parameters</partinfo>
 
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Revision as of 09:54, 31 August 2019


Xre

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 the res-xre locus from Photorhabdus luminescens and other bacterial species function as bona fide TA modules in Escherichia coli.

It was shown that a RES toxin encoded by the α‐proteobacterium, Sinorhizobium meliloti, is toxic when expressed in Escherichia coli and can be counteracted by its cognate Xre antitoxin.

RES toxin from several bacterial species reversibly inhibits cell growth in E. coli, and that expression of the Xre antitoxin neutralizes this toxicity. However, the cellular function of the RES toxin when activated, as well as structure and the antitoxic mechanism of the Xre antitoxin, remain unknown.

Biology

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


Characterisation

References

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.

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