Cloning and expression of Chlamydia trachomatis OmcBc gene and antigenicity analysis of the protein / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1558-1561, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-336143
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the antigenicity of recombinant Chlamydia trachomatis (Ct) OmcBc protein and search for the new target for early diagnosis of Chlamydia infection and Chlamydia vaccine development.</p><p><b>METHODS</b>The C fragment of OmcB encoding the amino acids from T270 to T553 was amplified from Chlamydia serovar D genomic DNA. The pGEX-6p-Ct OmcBc expression plasmid was constructed and transformed into E.coli XL-1blue. The expression of recombinant Ct OmcBc protein was induced by IPTG. Serum samples were collected from 120 patients with urogenital Chlamydia infection. The antiserum samples were collected from 7 New Zealand white rabbits and 5 Balb/C mice immunized subcutaneously and intraperitoneally with Ct serovar D inactivated EB, respectively, and from 9 Balb/C mice intranasally infected with Ct serovar D live EB. The anti-Chlamydia specific antibody were titrated by an immunofluorescence assay (IFA). The reactivity of the recombinant OmcBc protein with all the above antisera was detected by ELISA.</p><p><b>RESULTS</b>The pGEX-6p-Ct OmcBc expression plasmid was successfully constructed. DNA sequencing showed that the inserted OmcBc was about 852 bp, encoding a protein with 284 amino acids. The expression of the recombinant GST-OmcBc was induced by IPTG, producing a fusion protein with a molecular weight of about 57 kD. The titer of the specific antibodies to Chlamydia in all the antisera was high. ELISA results showed strong reactivities of the recombinant GST-OmcBc fusion protein with all the above antisera.</p><p><b>CONCLUSIONS</b>OmcBc protein is an immunodominant protein of Chlamydia. The recombinant GST-OmcBc with strong antigenicity may provide a basis for further study of early diagnosis of chlamydia infection and development of Chlamydia vaccine.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Proteínas da Membrana Bacteriana Externa
/
Sangue
/
Chlamydia trachomatis
/
Clonagem Molecular
/
Alergia e Imunologia
/
Genes Bacterianos
/
Genética
/
Soros Imunes
/
Metabolismo
Tipo de estudo:
Estudo de rastreamento
Limite:
Animais
/
Humanos
Idioma:
Chinês
Revista:
Journal of Southern Medical University
Ano de publicação:
2010
Tipo de documento:
Artigo
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