Polymorphisms in Chlamydia trachomatis tryptophan synthase genes differentiate between genital and ocular isolates
J. Clin. Invest. Harlan D. Caldwell, et al. 111:1757 doi:10.1172/JCI17993 [
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Figure 3RT-PCR quantitation of
trpBA and
trpR mRNA isolated from HeLa cell monolayers infected with
C. trachomatis ocular (serovar A 2497) and genital (serovars E IU823) clinical isolates cultured in the presence of IFN-γ. Culture and chlamydiae infection conditions were as described in the legend to Figure
2. Infected HeLa cells were cultured in the presence of complete DMEM-10 (+Trp), complete DMEM-10 plus 5 ng/ml IFN-γ (+IFN), complete DMEM-10 plus IFN-γ and 100 μM indole (IFN+Ind), and complete DMEM-10 plus IFN-γ and supplemented with 1 g/l tryptophan (IFN+Trp). RNA was isolated 36 hours after infection. The RNA was reverse transcribed using random hexamer primers, and the cDNA was used for quantitative PCR amplifications with primers specific for
trpBA or
trpR. Quantitative PCR reactions were carried out using the LightCycler and SYBR Green I as the fluorophore. See Methods for details.
