EXPRESSION OF A PLASMID GENE OF CHLAMYDIA-TRACHOMATIS ENCODING A NOVEL 28-KDA ANTIGEN

被引:49
作者
COMANDUCCI, M
CEVENINI, R
MORONI, A
GIULIANI, MM
RICCI, S
SCARLATO, V
RATTI, G
机构
[1] IMMUNOBIOL RES INST SIENA, VIA FIORENTINA 1, I-53100 SIENA, ITALY
[2] ST ORSOLA HOSP, INST MICROBIOL, I-40138 BOLOGNA, ITALY
来源
JOURNAL OF GENERAL MICROBIOLOGY | 1993年 / 139卷
关键词
D O I
10.1099/00221287-139-5-1083
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Plasmid pCT is present in essentially all isolates of Chlamydia trachomatis and may encode factors important for survival in the natural environment. However, no pCT-associated phenotype has been described so far. With the purpose of investigating the possibility of a role of pCT in C. trachomatis pathogenicity, we examined the expression of an ORF (ORF3), potentially encoding a 28 kDa polypeptide (pgp3). Analysis of RNA extracted from chlamydia-infected Vero cells detected ORF3-specific transcripts, from 20 h post-infection onwards, mainly as discrete RNA species of 1390 nucleotides comprising the downstream ORF4 sequence. ORF3 DNA was cloned and expressed in Escherichia coli as a 39 kDa fusion protein (MS2/pgp3). Antibodies raised against purified MS2/pgp3, specifically recognized a 28 kDa protein on Western blots of protein from purified chlamydial elementary bodies (EBs). The same antibodies detected chlamydial inclusions in methanol-fixed infected cells by immunofluorescence. Western blot analysis of EBs extracted with 2 % Sarkosyl, showed that a large proportion of the 28 kDa antigen is associated with the detergent-insoluble ('membrane') fraction. Antibodies recognizing pgp3 epitopes were detected in sera from patients with chlamydial infections, but not in sero-negative control sera. The findings support the hy thesis that pCT may provide a function related to chlamydial cell physiology.
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页码:1083 / 1092
页数:10
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