Part:BBa_K4047016:Experience

The gentamicin resistance worked as expected, in which some transformants were able to grow on a gentamicin-containing agar plate. This antibiotic resistance marker was used in the plasmid BBa_K4047039. The insertion of gentamicin indicated unsuccessful deletion of the target gene glpX, allowing us to distinguish double versus single crossovers due to the presence of gentamicin in the plasmid backbone.

This part was successfully used in the 2021 MiamiU_OH team project, with a picture shown below of the resulting growth, which was able to be compared to a cloned kanamycin plate to identify those successful deletion mutants that could only grow on kanamycin.

Colony growth on a gentamicin-containing BG11 plate in the MiamiU_OH 2021 project, indicating integration of the plasmid backbone and functional expression of this resistance gene.

Applications of BBa_K4047016

This part can be used in plasmids as a selective marker, indicating which colonies have integrated the entire plasmid and not deleted expression of the target gene.

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