Identification, Bioinformatics Analyses, and Expression of Immunoreactive Antigens of Mycoplasma haemofelis

Add to your list(s) Download to your calendar using vCal

• Professor Rose Raskin & Joanne B. Messick, Dept of Pathology College of Veterinary Medicine Purdue University, Indianna USA
• Wednesday 15 June 2011, 16:30-17:30
• Lecture Theatre 1, Department of Veterinary Medicine.

If you have a question about this talk, please contact Suzy Blows.

Mycoplasma haemofelis infection frequently causes anemia in cats. Despite an intense immune response and/or antibiotic treatment, cats often remain asymptomatic carriers following infection. Our hypothesis is that detection of antibodies to M. haemofelis is a sensitive approach for identifying infected cats, particularly carriers. To date, no immunoassay has been developed. This is due largely to the inability to culture M. haemofelis in vitro; hence a source of antigen is not readily available. The objective of this study was to identify, express and purify immunogenic proteins of M. haemofelis. To accomplish this, two whole-genomic expression libraries were created in the Lambda Zap®II vector and immunoscreened with pre-immune plasma, plasma from SPF cats, and pooled acute and convalescent-phase plasma from experimentally infected cats. The inserts from 21 immunoreactive clones were sequenced, resulting in the identification of 60 genes coding for putative proteins necessary for diverse cellular functions, along with several novel genes of M. haemofelis. Fragments of selected genes based on bioinformatic analyses were PCR amplified, cloned into a high-level protein expression system and subsequently expressed in Escherichia coli as a His6-fusion protein. The recombinant fusion proteins of M. haemofelis were purified and evaluated as an antigen in Western blot to verify the findings of previous immunoscreening. Together with bioinformatics analyses of individual genes, this approach provided several putative candidate antigens. Five antigens of M. haemofelis were reactive by Western blot against the immune plasma and negative against non-immune plasma; these antigens might be useful serologic or even vaccine targets.

This talk is part of the Departmental Seminar Programme, Department of Veterinary Medicine series.

This talk is included in these lists:

• All Talks (aka the CURE list)
• Cambridge Immunology
• Cambridge Infectious Disease
• Cambridge Infectious Diseases
• Departmental Seminar Programme, Department of Veterinary Medicine
• Lecture Theatre 1, Department of Veterinary Medicine
• Vet School Seminars

Note that ex-directory lists are not shown.