Difference between revisions of "Part:BBa K3198002"
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<partinfo>BBa_K3198002 short</partinfo> | <partinfo>BBa_K3198002 short</partinfo> | ||
+ | <!-- This part contains the toxin component of a type II toxin-antitoxin (TA) system. HicA is a probable translation-independent mRNA interferase.<br><br> --> | ||
+ | <br><br> | ||
+ | |||
+ | <span class='h3bb'>Sequence and Features</span> | ||
+ | <partinfo>BBa_K3198002 SequenceAndFeatures</partinfo> | ||
+ | |||
+ | ===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=== |
− | === | + | This part is from the res-xre locus from Photorhabdus luminescens and other bacterial species. |
+ | |||
+ | |||
+ | ===Characterisation=== | ||
+ | <!-- 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. | ||
+ | <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. | ||
+ | <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. | ||
+ | <br><br>Figure 1: Growth curve of control <i>MG1655</i> unaffected by IPTG | ||
+ | <br><br>Figure 2: Growth curve of <i>MG1655</i> transformed with HicA-containing plasmid | ||
+ | <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. | ||
+ | <br><br>Figure 3: Total GFP curve of HicA-plasmid containing cells with different IPTG induction (0M, 100μM, 500μM and 2mM) | ||
+ | <br><br>In summary, we believe that (BBa_K3198000) is a new BioBrick capable of causing cell growth arrest and suppressing protein expression. --> | ||
+ | |||
+ | |||
+ | ===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 | ||
+ | <br><br> | ||
+ | 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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<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
− | <partinfo> | + | <partinfo>BBa_K3198001 parameters</partinfo> |
<!-- --> | <!-- --> |
Revision as of 09:51, 31 August 2019
RES
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.