Difference between revisions of "Part:BBa K3796203"

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MazF is an mRNA interferase which cleaves mRNAs at a specific site, and <i>ndoA</i>, also called <i>ydcE</i> or <i>mazF-bs</i> shows a similar activity according to references given below. ndoA belongs to a type Ⅱ toxin-antitoxin system in Bacillus subtilis. There is great possibility that no antitoxin exists in C. glutamicum to inhibit EndoA encoded by ndoA. So it can be used to kill engineered <i>C. glutamicum</i> at a proper time.
 
MazF is an mRNA interferase which cleaves mRNAs at a specific site, and <i>ndoA</i>, also called <i>ydcE</i> or <i>mazF-bs</i> shows a similar activity according to references given below. ndoA belongs to a type Ⅱ toxin-antitoxin system in Bacillus subtilis. There is great possibility that no antitoxin exists in C. glutamicum to inhibit EndoA encoded by ndoA. So it can be used to kill engineered <i>C. glutamicum</i> at a proper time.
 
===Characterization===
 
===Characterization===
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This year, our team designed a kill switch in <i>Corynebacterium glutamicum </i> using <i>ndoA</i> from <i>Bacillus subtilis</i> subsp. subtilis str. 168. We aimed to test if the overexpression of <i>ndoA</i> in <i>Corynebacterium glutamicum</i> can kill the engineered bacteria effectively, and documented our works and failures in iGEM Parts to inspire future iGEM teams for kill switch design.
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This year, our team designed a kill switch in <i>C. glutamicum</i> using <i>ndoA</i> from <i>Bacillus subtilis</i> subsp. subtilis str. 168 under the control of PgsiB (alkali inducible promoter) and Patp2 (alkali inducible promoter). We aimed to test if the over-expression of <i>ndoA</i> in <i>C. glutamicum</i> can kill the engineered bacteria effectively, and documented our works and failures in iGEM Parts to inspire future iGEM teams for kill switch design.

Revision as of 16:19, 11 October 2021


ndoA (endoribonuclease toxin in Bacillus subtilis)

This coding region ndoA(also mazF, ydcE) encodes endoribonuclease EndoA, a toxic component of a type II toxin-antitoxin (TA) system. It encodes endoribonuclease toxin in Bacillus subtilis that is a UACAU‐specific mRNA interferase, cleavaging after the first U. Previous research has proved that its overexpression is toxic for cell growth and it functions as normal in Escherichia coli.


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 and Biology

MazF is an mRNA interferase which cleaves mRNAs at a specific site, and ndoA, also called ydcE or mazF-bs shows a similar activity according to references given below. ndoA belongs to a type Ⅱ toxin-antitoxin system in Bacillus subtilis. There is great possibility that no antitoxin exists in C. glutamicum to inhibit EndoA encoded by ndoA. So it can be used to kill engineered C. glutamicum at a proper time.

Characterization

This year, our team designed a kill switch in C. glutamicum using ndoA from Bacillus subtilis subsp. subtilis str. 168 under the control of PgsiB (alkali inducible promoter) and Patp2 (alkali inducible promoter). We aimed to test if the over-expression of ndoA in C. glutamicum can kill the engineered bacteria effectively, and documented our works and failures in iGEM Parts to inspire future iGEM teams for kill switch design.