Difference between revisions of "Part:BBa K3447108"
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[[Image: T--Jilin_China--Results--14.jpg|center|thumb|600px|<b>Fig. 1 Kan<sup>R</sup> has resistance of kanamycin.</b> (A) Digestion and electrophoresis of Kan<sup>R</sup>. (B) Functional verification of Kan<sup>R</sup>. <i>E. coli</i> DH5α was transformed with the designed plasmid, cultured overnight, and diluted OD<sub>600</sub> to 0.02. Measurement at the indicated time with 0.1% kanamycin added. The experiment was performed three times in triplicate. *, P < 0.05 compared with respective control. (C) Zone of inhibition test. 50 μL suspension was spread over the face of LB agar plate, on which a hole was bored for 0.1% kanamycin added. Incubate at 37℃ for 12 hrs. The plasmid was transformed into <i>E. coli</i> DH5α, cultured overnight, and diluted OD600 to 0.02.]]<br> | [[Image: T--Jilin_China--Results--14.jpg|center|thumb|600px|<b>Fig. 1 Kan<sup>R</sup> has resistance of kanamycin.</b> (A) Digestion and electrophoresis of Kan<sup>R</sup>. (B) Functional verification of Kan<sup>R</sup>. <i>E. coli</i> DH5α was transformed with the designed plasmid, cultured overnight, and diluted OD<sub>600</sub> to 0.02. Measurement at the indicated time with 0.1% kanamycin added. The experiment was performed three times in triplicate. *, P < 0.05 compared with respective control. (C) Zone of inhibition test. 50 μL suspension was spread over the face of LB agar plate, on which a hole was bored for 0.1% kanamycin added. Incubate at 37℃ for 12 hrs. The plasmid was transformed into <i>E. coli</i> DH5α, cultured overnight, and diluted OD600 to 0.02.]]<br> | ||
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+ | ==<b>Design</b>== | ||
+ | ===Design Notes=== | ||
+ | We added some synonymous mutations to avoid part rules.<br> | ||
+ | |||
+ | |||
+ | |||
+ | ===Source=== | ||
+ | |||
+ | We found this sequence data in the previous iGEM teams (Glasgow 2017 and <partinfo>BBa_I0500</partinfo>) and in GenBank.<br> | ||
+ | |||
+ | ===References=== | ||
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Revision as of 09:51, 27 October 2020
Expression of kanamycin resistance
This is a part with kanamycin resistance gene (BBa_K3447003).
Usage and Biology
It is well-known that kanamycin has the ability to inhibit the growth of bacteria, so we expect that with the expression of KanR gene, our engineered bacteria would be able to survive under the attack of kanamycin.
Characterization
To verify the construction of KanR, the digestion and agarose gel electrophoresis were performed by a standard protocol (Fig. 1A). Compared with the control group, bacteria constructed with KanR show more than 10 times the resistance to kanamycin (Fig. 1B). Zone of inhibition test also shows that our KanR works well with 0.1% kanamycin added (Fig. 1C).
Design
Design Notes
We added some synonymous mutations to avoid part rules.
Source
We found this sequence data in the previous iGEM teams (Glasgow 2017 and BBa_I0500) and in GenBank.
References
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 26
Illegal NheI site found at 49 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 110
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1
Illegal BsaI.rc site found at 1039