Dana Furst

Defense Date

7-28-2005

Graduation Date

Summer 2005

Availability

Immediate Access

Submission Type

thesis

Degree Name

MS

Department

Biological Sciences

Committee Chair

Peter A. Castric

Committee Member

Kyle W. Selcer

Committee Member

Nancy Trun

Keywords

Biofilm, Gram-negative Bacteria, Microbial Physiology, Twitching Motility

Abstract

The presence of Pseudomonas aeruginosa 1244 pili on the bacterial surface is dependent upon the growth environment. Quantitative western immunoblot analysis showed that an agar plate promoted surface expression of pili, whereas bacterial cells grown in liquid broth lacked similar appendages. Hyperexpression of the pilA gene increased piliation in broth cells but not to the extent of plate cells. Real-time RT-PCR indicated that the pilA gene has the potential to be regulated posttranscriptionally, regardless of the growth environment.

The P. aeruginosa 1244 pilin glycan and lipopolysaccharide O-antigen are synthesized via the same biosynthetic pathway. Rabbits immunized with LPS-free pili produced antibodies that recognized both pili and LPS. Serum from these animals was analyzed using ELISA assays, and the pilin glycan was found to be an important immunogenic epitope directing strong responses against both the glycan and O-antigen. This could prove useful in P. aeruginosa vaccine generation.

Format

PDF

Language

English

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