Difference between revisions of "Part:BBa K2120004"
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Arac-pBAD is an arabinose induced promoter, mazF is a toxin gene. This part is functioned as a killer. | Arac-pBAD is an arabinose induced promoter, mazF is a toxin gene. This part is functioned as a killer. | ||
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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), | ||
+ | [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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE 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.
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).