Blackburn, Paul Edward
Characterisation of the virulence-related, outer-membrane proteins of B. pertussis.
PhD thesis, University of Glasgow.
Full text available as:
At the beginning of this study, knowledge of autotransporter proteins was in its infancy and the B. pertussis proteins pertactin, BrkA and tracheal colonisation factor (Tcf) had not been recognised as members of this family. However, the sequence similarity between these Bordetella proteins and a number of other proteins from different genera soon became apparent and the Bordetella proteins were also included in the autotransporter protein family. It was the similarity between pertactin and other B. pertussis proteins that, due to a mis-primed PCR amplification, led to the preliminary data for this project. A B. pertussis genomic library was screened, using this amplicon s a Southern blot probe, and a cosmid was identified and subcloned. Analysis of the sequence led to the identification of a larger open reading frame which had not been previously recognised. The cosmid did not contain the entire open reading frame and a further 200 bp was obtained using an adapter-linked single specific primer-PCR (SSP-PCR). An open reading frame was predicted and the amino acid similarity strongly suggests that this gene product represents another member of the autotransporter family. This protein was named Bap-5 as the fifth member of the Bordetella autotransporter protein family. Motif analysis revealed an RGD integrin-binding motif and an SGSG glycosaminoglycan-binding motif, which implicates this protein as a putative adhesin.
Although not begun at the start of this study, the B. pertussis genome sequence was near completion during the final stages of this work and a sequence identical to bap-5 was located in the sequence data.
Actions (login required)