Difference between revisions of "Part:BBa K2120004"
 
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<partinfo>BBa_K2120004 short</partinfo>
 
<partinfo>BBa_K2120004 short</partinfo>
  
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This is a part of 2016 BIT-China iGEM Team to verify the function and character of the mazF toxin gene. And it also the verification circuit of the killer device in the project.
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Arac-pBAD is an arabinose induced promoter, mazF is a toxin gene. This part is functioned as a killer.  
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We got the circuit by using one-step mutation from araC+PBAD+B0032+mazF [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120002 BBa_K2120002]. And we constructed the plasmid containing araC+PBAD+B0031+mazF circuits, and transformed the plasmid into top10. After transformation, we measured the OD600 to draw the growth curve and observed the function of toxin protein. Besides the bacteria contained the toxin gene, the empty pSB1C3 vector was the control. Add arabinose or not, there was another comparison. We add 10% arabinose 50 uL into 50 mL LB culture medium when the OD600 is 0.6 (the log phase). We measured the OD600 every hour until the bacteria reached the stationary phase.
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[[File:BIT-CHINA-PARTS-KILLER-8.jpg|600px|thumb|center|Fig. Compared with control groups, the difference value of OD600 showed the lethal efficiency of the circuit]]
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According to the results, only the bacteria containing the plasmid with toxin gene and induced by arabinose can be inhibited and killed.
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For more results, please see:
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[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120002 BBa_K2120002](araC+PBAD+B0032+mazF),
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[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120003 BBa_K2120003](araC+PBAD+B0032+hokD),
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[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120005 BBa_K2120005](araC+PBAD+B0031+hokD),
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[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120006 BBa_K2120006](araC+PBAD+B0034+mazF).
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<partinfo>BBa_K2120004 parameters</partinfo>
 
<partinfo>BBa_K2120004 parameters</partinfo>
 
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===Functional Parameters===
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 +
This is a part of 2016 BIT-China iGEM Team to verify the function and character of the mazF toxin gene. And it also the verification circuit of the killer device in the project.
 +
We got the circuit by using one-step mutation from araC+PBAD+B0032+mazF [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120002 BBa_K2120002]. And we constructed the plasmid containing araC+PBAD+B0031+mazF circuits, and transformed the plasmid into top10. After transformation, we measured the OD600 to draw the growth curve and observed the function of toxin protein. Besides the bacteria contained the toxin gene, the empty pSB1C3 vector was the control. Add arabinose or not, there was another comparison. We add 10% arabinose 50 uL into 50 mL LB culture medium when the OD600 is 0.6 (the log phase). We measured the OD600 every hour until the bacteria reached the stationary phase.
 +
[[File:BIT-CHINA-PARTS-KILLER-8change.png|600px|thumb|center|Fig. Compared with control groups, the difference value of OD600 showed the lethal efficiency of the circuit]]
 +
According to the results, only the bacteria containing the plasmid with toxin gene and induced by arabinose can be inhibited and killed.
 +
For more results, please see:
 +
[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120002 BBa_K2120002](araC+PBAD+B0032+mazF),
 +
[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120003 BBa_K2120003](araC+PBAD+B0032+hokD),
 +
[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120005 BBa_K2120005](araC+PBAD+B0031+hokD),
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[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120006 BBa_K2120006](araC+PBAD+B0034+mazF).

Latest revision as of 20:57, 19 October 2016


araC+PBAD+B0031+mazF

Arac-pBAD is an arabinose induced promoter, mazF is a toxin gene. This part is functioned as a killer.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Functional Parameters

This is a part of 2016 BIT-China iGEM Team to verify the function and character of the mazF toxin gene. And it also the verification circuit of the killer device in the project. We got the circuit by using one-step mutation from araC+PBAD+B0032+mazF BBa_K2120002. And we constructed the plasmid containing araC+PBAD+B0031+mazF circuits, and transformed the plasmid into top10. After transformation, we measured the OD600 to draw the growth curve and observed the function of toxin protein. Besides the bacteria contained the toxin gene, the empty pSB1C3 vector was the control. Add arabinose or not, there was another comparison. We add 10% arabinose 50 uL into 50 mL LB culture medium when the OD600 is 0.6 (the log phase). We measured the OD600 every hour until the bacteria reached the stationary phase.

Fig. Compared with control groups, the difference value of OD600 showed the lethal efficiency of the circuit

According to the results, only the bacteria containing the plasmid with toxin gene and induced by arabinose can be inhibited and killed. For more results, please see: BBa_K2120002(araC+PBAD+B0032+mazF), BBa_K2120003(araC+PBAD+B0032+hokD), BBa_K2120005(araC+PBAD+B0031+hokD), BBa_K2120006(araC+PBAD+B0034+mazF).