Difference between revisions of "Part:BBa K733004"
 
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<partinfo>BBa_K733004 short</partinfo>
 
<partinfo>BBa_K733004 short</partinfo>
  
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ydcE which also is named as ndoA, encoding an RNase, namely EndoA. Being in the same RNase family with MazF/ChpAK/PemK, YdcE - the endoribonuclease ydcE encodes - can inactivate cellular mRNAs by cleaving them at specific but frequently occurring sites (i.e. UAUAAU↓AC). Thus, overexpression of EndoA can lead to programmed cell death or unhealthy conditions, depending on the amount of EndoA. Note that ydcE(ndoA) is demonstrated functional both in E. coli and B. subtilis.  (Pellegrini et al., 2005)
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''ydcE'' which is also named as ''ndoA'', encoding an RNase, namely EndoA. Being in the same RNase family with MazF/ChpAK/PemK, YdcE - the endoribonuclease ''ydcE'' encodes - can inactivate cellular mRNAs by cleaving them at specific but frequently occurring sites (i.e. UAUAAU↓AC). Thus, overexpression of EndoA can lead to programmed cell death or unhealthy conditions, depending on the amount of EndoA. Note that ''ydcE(ndoA)'' is demonstrated functional both in'' E. coli'' and ''B. subtilis''.  (Pellegrini et al., 2005)
  
 
[[Image:YdcE.png]]
 
[[Image:YdcE.png]]
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<h3>Characterization by UCAS iGEM 2016 team</h3>
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This summer we used the ydcD and ydcE genes to build a kill-switch.
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By measuring the growth curve of E. coli expressing toxin, we showed that toxin EndoA can effectively impede the growth and cleavage of E. coli.
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We also characterized antitoxin ydcD, and documented the interaction between toxin and antitoxin on the page:
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[https://parts.igem.org/Part:BBa_K733003 BBa_K733003]
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[[file:T--UCAS--ydcE1.png|500px]]
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From the graph above we see that toxin is induced by graded anhydrotetracycline. It shows significant toxicity when induced 1.25 and 2.5µg/mL of aTc is added.
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We also performed morphological analysis of E. coli expressing EndoA by SEM, and we found that the bacteria change from rod-like shape to sphere-like shape.
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[[Image:T--UCAS--1204sem.png|500px]]
  
 
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==Reference==
 
==Reference==
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Pellegrini O, Mathy N, Gogos A, Shapiro L, and Condon C. "The Bacillus subtilis ydcDE operon encodes an endoribonuclease of the MazF/PemK family and its inhibitor.." Molecular microbiology. 56.5 (2005): 1139-1148. Print.
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Pellegrini O, Mathy N, Gogos A, Shapiro L, and Condon C. "The ''Bacillus subtilis'' ''ydcDE'' operon encodes an endoribonuclease of the MazF/PemK family and its inhibitor.." ''Molecular microbiology''. 56.5 (2005): 1139-1148. Print.

Latest revision as of 02:31, 20 October 2016

RBS+ydcE

ydcE which is also named as ndoA, encoding an RNase, namely EndoA. Being in the same RNase family with MazF/ChpAK/PemK, YdcE - the endoribonuclease ydcE encodes - can inactivate cellular mRNAs by cleaving them at specific but frequently occurring sites (i.e. UAUAAU↓AC). Thus, overexpression of EndoA can lead to programmed cell death or unhealthy conditions, depending on the amount of EndoA. Note that ydcE(ndoA) is demonstrated functional both in E. coli and B. subtilis. (Pellegrini et al., 2005)

YdcE.png

Characterization by UCAS iGEM 2016 team

This summer we used the ydcD and ydcE genes to build a kill-switch. By measuring the growth curve of E. coli expressing toxin, we showed that toxin EndoA can effectively impede the growth and cleavage of E. coli.

We also characterized antitoxin ydcD, and documented the interaction between toxin and antitoxin on the page: BBa_K733003

T--UCAS--ydcE1.png

From the graph above we see that toxin is induced by graded anhydrotetracycline. It shows significant toxicity when induced 1.25 and 2.5µg/mL of aTc is added.

We also performed morphological analysis of E. coli expressing EndoA by SEM, and we found that the bacteria change from rod-like shape to sphere-like shape.

File:T--UCAS--1204sem.png

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]


Reference

Pellegrini O, Mathy N, Gogos A, Shapiro L, and Condon C. "The Bacillus subtilis ydcDE operon encodes an endoribonuclease of the MazF/PemK family and its inhibitor.." Molecular microbiology. 56.5 (2005): 1139-1148. Print.