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Mapping of Chlamydia trachomatis proteins by Immobiline‐polyacrylamide two‐dimensional electrophoresis: Spot identification by N ‐terminal sequencing and immunoblotting
Author(s) -
Bini Luca,
SanchezCampillo Maria,
Santucci Annalisa,
Magi Barbara,
Marzocchi Barbara,
Comanducci Maurizio,
Christiansen Gunna,
Birkelund Sevend,
Cevenini Roberto,
Vretou Evangelia,
Ratti Giulio,
Pallini Vitaliano
Publication year - 1996
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.1150170130
Subject(s) - biology , chlamydia trachomatis , bacterial outer membrane , microbiology and biotechnology , gel electrophoresis , ribosomal protein , polyacrylamide gel electrophoresis , membrane protein , chlamydiaceae , biochemistry , gene , escherichia coli , ribosome , membrane , enzyme , rna , immunology
Proteins from purified elementary bodies of Chlamydia trachomatis were separated by two‐dimensional gel electrophoresis on nonlinear wide‐range immobilized pH gradients in the first dimension and polyacrylamide gradient gels in the second dimension. The maps obtained with this system are highly reproducible and resolve ca. 600 spots. By using immunoblot analysis with specific antibodies and/or N ‐terminal amino acid sequencing, we established the positions of a number of described chlamydial proteins, such as the major outer membrane protein (MOMP) the 60 kDa cystein‐rich outer membrane protein (OMP2), the DnaK‐like, GroEL‐like, and macrophage infectivity potentiator (MIP)‐like proteins, the plasmid‐encoded pgp3 protein, two ribosomal proteins (S1 and L7/L12), and the protein‐elongation factor EF‐Tu. Other proteins, for which gene assignment was not possible, have been identified by three parameters ( M r , p I and N ‐terminal sequence). This work provides a preliminary basis for a future and progressive compilation of a genome‐linked database of chlamydial proteins.