Analyzing the Function of NucS in M. smegmatis

Importance

Antimicrobial resistance is one of the top ten threats to global health. Antimicrobial resistance in bacteria is often caused by mutations. Many bacteria have innate systems to prevent mutations. These systems involve groups of proteins working together to identify and get rid of mutations. NucS is one such mutation-preventing protein that we hope to learn more about in this project.

Grant Proposal

Project Images

A simplified image of the pEW2 plasmid. The <q>5</q> represents the sequence coding for the five amino acids at the end of the NucS protein.

A simplified image of the pEW2 plasmid. The 5 represents the sequence coding for the five amino acids at the end of the NucS protein.

A gel of pEW2-2 and pEW2-3 for junctions present in the correct plasmid.

A gel running plasmid candidates for junctions present in the intended plasmid. pEW2-2 and pEW2-3 had the intended junctions, as seen by the two bands present in the fourth and fifth lanes where they were run.

Some rifampicin plates.

Some rifampicin plates we used to determine the frequency of mutation of each strain of bacteria.

A rifampicin plate of a strain containing a deletion in nucS with 172 mutagenic colonies.

A strain of M. smegmatis that had a deletion in its nucS gene plated on a rifampicin plate. We counted 172 mutagenic colonies.

Project Notes

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