Difference between revisions of "Part:BBa K3447108"

Latest revision as of 10:24, 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 Kan^R gene, our engineered bacteria would be able to survive under the attack of kanamycin.

Characterization

To verify the construction of Kan^R, the digestion and agarose gel electrophoresis were performed by a standard protocol (Fig. 1A). Compared with the control group, bacteria constructed with Kan^R show more than 10 times the resistance to kanamycin (Fig. 1B). Zone of inhibition test also shows that our Kan^R works well with 0.1% kanamycin added (Fig. 1C).

Fig. 1 Kan^R has resistance of kanamycin. (A) Digestion and electrophoresis of Kan^R. (B) Functional verification of Kan^R. E. coli DH5α was transformed with the designed plasmid, cultured overnight, and diluted OD₆₀₀ 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 E. coli DH5α, cultured overnight, and diluted OD600 to 0.02.

Design

Design Notes

We added some synonymous mutations to avoid part rules.

Source

We found this sequence data in GenBank.

References

Sequence and Features