Abstract

Francisella tularensis is a pathogenic gram-negative bacterium and causes a pneumonic type of often fatal disease called Tularaemia. F. tularensis can infect a large number of mammalian species including humans. It is a fastidious, facultative intracellular bacterium, which requires cysteine for growth. Tularemia is treated with anti-microbials, for example, aminoglycosides, antibiotic  medications, or fluoroquinolones. The objective of the assignment is to clone a PCR target gene (TUL4) of F.tularensis in E.coli. PCR was performed using genomic DNA of F. tularensis as a template. After PCR, the amplicon was purified and cloned into a cloning vector. Positive clones were selected based on colony PCR screening. Recombinant plasmids from the positive E. coli clones were purified and kept for subsequent downstream applications.