Cloning and characterization of a Chlamydia trachomatis L3 DNA fragment that codes for an antigenic region of the major outer membrane protein and specifically hybridizes to the C- and C-related-complex serovars.
Chlamydia trachomatis L3 DNA was cloned and expressed in lambda gt11. A recombinant plaque that expressed an antigen that reacted with rabbit polyclonal antichlamydial L3 serum and with two monoclonal antibodies specific for serovars L3 and I was selected from this Chlamydia genomic library. The beta-galactosidase Chlamydia fusion protein was purified by immunoaffinity chromatography and injected into mice to produce monoclonal antibodies. These monoclonal antibodies reacted by Western (immuno-) blot with both the fusion protein and the major outer membrane protein from purified L3 elementary bodies. The chlamydial DNA fragment was shown by DNA sequence analysis to be 168 base pairs in length and to correspond to the constant regions 1 and 2 and the variable segment 1 of the major outer membrane protein gene. The recombinant chlamydial DNA fragment hybridized under stringent conditions by Southern and dot blot analysis exclusively with the DNA from the C- and C-related-complex C. trachomatis serovars.
Carlson
E J
EJ
Department of Pathology, University of California, Irvine 92717.
Peterson
E M
EM
de la Maza
L M
LM
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002692:Chlamydia trachomatis / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000302:isolation & purification*; D005260:Female; D005798:Genes, Bacterial; D015151:Immunoblotting; D008565:Membrane Proteins / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D012703:Serotyping; D001616:beta-Galactosidase / Q000235:genetics / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase