Klebsiella pneumoniae KPPR1 Insertion Site Sequencing (InSeq) in Lung Infection

Klebsiella pneumoniae is a common cause of hospital-acquired pneumonia and an urgent threat to public health due to its increasing antibiotic resistance. The goal of this study is to identify K. pneumoniae genes that contribute to its fitness during lung infection. A pool of transposon insertion mutants was constructed and used to infect mouse lungs. Genomic DNA was extracted from the inoculum and K. pneumoniae recovered from the lungs of infected mice, and fragments of DNA at the junction between the transposon insertion and the surrounding genome were generated. These fragments were sequenced by Illumina 50 bp single end sequencing and used to map and count the frequency of each transposon insertion in the inoculum and lung samples. The ratio of inoculum/lung counts was used to measure the relative fitness contribution of the disrupted gene during lung infection.