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GENETIC CHARACTERIZATION OF THE, K2 SEROTYPE CAPSUL.E OF KLEBSIELLA PNEUMONIAE ATCC 43816 AND THE DEVELOPME~IT OF A BIOLUMINESCENT STRAIN Jacob Brandon Scott April 23, 2013 OBJECTIVE: Klebsiella pneumoniae is a Gram-negative enterobacterium that is a major cause of community-acquired and nosocomial infections. We performed a genetic characterization of K. pneumoniae strain ATCC 43816, which is a wellstudied strain with a K2 serotype capsular polysaccharide. To provide thl~ ability to monitor bacteria within a host, we will develop a bioluminescence stra n of K. pneumoniae by integrating the lux operon into the chromosome. METHODS: The ATCC 43816 capsule cluster sequencing was performed by closing gapE, in the Next Generation Sequencing by long range peR for template and primer walking. We constructed a bioluminescent K. pneumoniae by using allelit:: exchange strategies and subsequently characterized the bioluminescent strain JSKP001 in a macrophage infection model. RESULTS: We have sequerced and annotated the CPS cluster as a 23,804-base pair sequence and identified eight homologous genes conserved between all serotypes of Klebsiella. Using our

Genetic characterization of the K2 serotype capsule of Klebsiella pneumoniae ATCC 43816 and the development of a bioluminescent strain.