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Two‐dimensional electrophoretic analysis of the protein family at 90 kDa of abortifacient Chlamydia psittaci
Author(s) -
Giannikopoulou Panagiota,
Bini Luca,
Simitsek Phaedra D.,
Pallini Vitaliano,
Vretou Evangelia
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150181137
Subject(s) - chlamydia psittaci , molecular mass , antigen , recombinant dna , biology , microbiology and biotechnology , antibody , gene , monoclonal antibody , chlamydia , genetics , biochemistry , enzyme
Four major clusters, designated A, B, C and D, were distinguished in Western Blots by a monoclonal antibody specific for the “antigen family at 90 kDa” after two‐dimensional electrophoretic analysis on immobilized pH gradient of chlamydial elementary bodies of abortifacient C. psittaci . Clusters B, C, and D were closely related with molecular mass (kDa) p I values of 91.5/5.2–5.4, 90/5.0–5.2 and 90.5/5.6–5.8, respectively. Cluster A was larger, with molecular mass/p I of 104.7/5.1–5.3. Evidence for the antigenic relationship between cluster A and clusters B, C and D was further supported by immunological cross‐reaction with affinity‐purified antibodies from serum of ewes with chlamydial‐induced abortion. The experimental values obtained for size and p I of the four clusters correlated well with the calculated values from known sequences coded by multiple chlamydial genes. Direct evidence for the correspondence between the immunoreactive clusters B, C and D and the retrieved genes was provided by antibody binding experiments to recombinant polypeptides representing fragments of the deduced proteins. The 4‐member antigen family at 90/104 kDa is the first example of proteins coded by multiple genes within the genus Chlamydia.