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SUMMARY:Identification\, Bioinformatics Analyses\, and Expression of Immun
 oreactive Antigens of Mycoplasma haemofelis - Professor Rose Raskin &amp\;
  Joanne B. Messick\, Dept of Pathology College of Veterinary Medicine Purd
 ue University\, Indianna USA
DTSTART:20110615T153000Z
DTEND:20110615T163000Z
UID:TALK30583@talks.cam.ac.uk
CONTACT:Suzy Blows
DESCRIPTION:Mycoplasma haemofelis infection frequently causes anemia in ca
 ts. 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 immunoass
 ay 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 immunoge
 nic proteins of M. haemofelis. To accomplish this\, two whole-genomic expr
 ession libraries were created in the Lambda Zap®II vector and immunoscree
 ned with pre-immune plasma\, plasma from SPF cats\, and pooled acute and c
 onvalescent-phase plasma from experimentally infected cats.  The inserts f
 rom 21 immunoreactive clones were sequenced\, resulting in the identificat
 ion of 60 genes coding for putative proteins necessary for diverse cellula
 r functions\, along with several novel genes of M. haemofelis.  Fragments 
 of selected genes based on bioinformatic analyses were PCR amplified\, clo
 ned into a high-level protein expression system and subsequently expressed
  in Escherichia coli as a His6-fusion protein.  The recombinant fusion pro
 teins of M. haemofelis were purified and evaluated as an antigen in Wester
 n blot to verify the findings of previous immunoscreening. Together with b
 ioinformatics analyses of individual genes\, this approach provided severa
 l putative candidate antigens. Five antigens of M. haemofelis were reactiv
 e by Western blot against the immune plasma and negative against non-immun
 e plasma\; these antigens might be useful serologic or even vaccine target
 s. 
LOCATION:Lecture Theatre 1\, Department of Veterinary Medicine
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