Difference between revisions of "Part:BBa K3198001"
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===Characterisation===
 
===Characterisation===
 
Team NUS 2019 has added another new biobrick (insert ID) into iGEM repository this year. This biobrick was found to possess the ability to neutralize the effect of HicA( biobrick id) and therefore functions as an antitoxin. For this reason, team NUS 2019 used this biobrick as part of their sleep-wake module to control the growth of E. coli in their project.
 
Team NUS 2019 has added another new biobrick (insert ID) into iGEM repository this year. This biobrick was found to possess the ability to neutralize the effect of HicA( biobrick id) and therefore functions as an antitoxin. For this reason, team NUS 2019 used this biobrick as part of their sleep-wake module to control the growth of E. coli in their project.
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</br>(HicB ID) was placed under an arabinose-inducible promoter and various arabinose concentrations were used to determine their ability to resume cell growth native MG1655.  
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<br>(HicB ID) was placed under an arabinose-inducible promoter and various arabinose concentrations were used to determine their ability to resume cell growth native MG1655.  
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</br>Characterization of cells transformed with this plasmid was performed at 37°C for 12h continuously. Both toxin and antitoxin inducers were added together at 0h, with IPTG concentration fixed at 500uM with varying arabinose concentrations of 0M, 0.0266M, 0.133M and 0.266M.  
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<br>Characterization of cells transformed with this plasmid was performed at 37°C for 12h continuously. Both toxin and antitoxin inducers were added together at 0h, with IPTG concentration fixed at 500uM with varying arabinose concentrations of 0M, 0.0266M, 0.133M and 0.266M.  
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</br>After the addition of 500μM IPTG into cells co-transformed with (HicA ID) and (HicB ID) plasmid, it was shown that even at the lowest arabinose concentration of 0.0266M, the growth of the cells was able to restore to the same level as uninduced cells. On the other hand, cells only induced with 500μM IPTG did not managed to show growth resumption in the absence of arabinose. This demonstrated the function of (HicB ID) as an antitoxin capable of neutralising (HicA ID) effect on native MG1655 growth.  
+
<br>After the addition of 500μM IPTG into cells co-transformed with (HicA ID) and (HicB ID) plasmid, it was shown that even at the lowest arabinose concentration of 0.0266M, the growth of the cells was able to restore to the same level as uninduced cells. On the other hand, cells only induced with 500μM IPTG did not managed to show growth resumption in the absence of arabinose. This demonstrated the function of (HicB ID) as an antitoxin capable of neutralising (HicA ID) effect on native MG1655 growth.  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:08, 31 August 2019


HicB

This part contains the antitoxin component of a type II toxin-antitoxin (TA) system. It functions as an mRNA interferase antitoxin; overexpression prevents HicA-mediated cessation of cell growth and cell death. HicB functions as an mRNA interferase antitoxin. Expression of HicB (145 aa) prevented HicA-mediated inhibition of cell growth. HicB neutralizes HicA and therefore functions as an antitoxin. HicB could resuscitate cells inhibited by HicA.

Usage

HicB functions as an mRNA interferase antitoxin. Expression of HicB (145 aa) prevented HicA-mediated inhibition of cell growth. HicB neutralizes HicA and therefore functions as an antitoxin. HicB could resuscitate cells inhibited by HicA.


Biology

HicB is from hicAB locus of Escherichia coli K-12.


Characterisation

Team NUS 2019 has added another new biobrick (insert ID) into iGEM repository this year. This biobrick was found to possess the ability to neutralize the effect of HicA( biobrick id) and therefore functions as an antitoxin. For this reason, team NUS 2019 used this biobrick as part of their sleep-wake module to control the growth of E. coli in their project.
(HicB ID) was placed under an arabinose-inducible promoter and various arabinose concentrations were used to determine their ability to resume cell growth native MG1655.
Characterization of cells transformed with this plasmid was performed at 37°C for 12h continuously. Both toxin and antitoxin inducers were added together at 0h, with IPTG concentration fixed at 500uM with varying arabinose concentrations of 0M, 0.0266M, 0.133M and 0.266M.
After the addition of 500μM IPTG into cells co-transformed with (HicA ID) and (HicB ID) plasmid, it was shown that even at the lowest arabinose concentration of 0.0266M, the growth of the cells was able to restore to the same level as uninduced cells. On the other hand, cells only induced with 500μM IPTG did not managed to show growth resumption in the absence of arabinose. This demonstrated the function of (HicB ID) as an antitoxin capable of neutralising (HicA ID) effect on native MG1655 growth.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 48
    Illegal BsaI.rc site found at 88