Project 1: An IncP-1β plasmid present in Pseudomonas aeruginosa isolated from the Burns Unit of Queen Elizabeth Hospital, Birmingham AND Project 2: Acid detection by the EvgS/A two component system

Aggarwal, Nikhil (2015). Project 1: An IncP-1β plasmid present in Pseudomonas aeruginosa isolated from the Burns Unit of Queen Elizabeth Hospital, Birmingham AND Project 2: Acid detection by the EvgS/A two component system. University of Birmingham. M.Res.

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Abstract

Project 1: An IncP-1β plasmid was found to be present in 13 Pseudomonas aeruginosa strains isolated from the Burns Unit of the Queen Elizabeth Hospital, Birmingham and subjected to High Throughput Sequencing (HTS). The sequence of the plasmid was analyzed and it was found that the plasmid has a chlorite dismutase gene inserted in its transposon, in lieu of an antibiotic resistance gene. The plasmid was tagged with an antibiotic resistance gene and its stability was determined in different bacterial strains. Isogenic strain without the plasmid was created and the biofilm formation activity and susceptibility to commonly used antibiotics was determined for plasmid positive and plasmid negative strains.

Project 2: EvgS is a sensor kinase of the EvgS/A two-component system involved in detecting acidic pH. Although the exact mechanism of acid detection is unknown, it is likely that the His residues in the periplasmic domains of EvgS play a role. Site-directed mutagenesis was employed to determine if any of the eight His residues in the first periplasmic domain are involved in acid detection. Role of different free amino acids and KCl in the activity of EvgS was also determined. Finally, affinity chromatography was used to purify the periplasmic domains of EvgS.

Type of Work:

Thesis (Masters by Research > M.Res.)

Award Type:

Masters by Research > M.Res.

Supervisor(s):