A molecular study of putative pili structures in Enterococuus faecium

Infections caused by Enterococcus faecium have increased over the last few decades. In this thesis there has been an attempt to identify putative pili proteins in E. faecium. The putative genes were ligated into a pENTR vector; the insert was sequenced to check the orientation, sequence, length and ends. Then the putative genes were recombined into a pDEST vector for expression. The expression showed that the protein was in the insoluble fraction, and that it was a bit smaller than expected. The prevalence of pili genes was done by screening a selection of isolates by PCR and check for positive isolates on an E-gel. Pili 1:44 showed a correlation to isolates from hospital survey and hospital outbreaks. While pili 1:48 showed a correlation to isolates from blood and vancomycin resistant isolates. The pili genes were also correlated to each other. The results indicate that the pili genes are expressed under different temperature conditions.