2012 Fiscal Year Final Research Report
Mycobacterial PE_PGRS62 gene is a virulence factor during tuberculosis.
| Project/Area Number |
22590411
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Research Institute, International Medical Center of Japan |
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Principal Investigator |
KIRIKAE Teruo 独立行政法人国立国際医療研究センター, 研究所, 部長 (50192563)
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| Co-Investigator(Kenkyū-buntansha) |
FUNATOGAWA Keiji (70536896)
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| Project Period (FY) |
2010 – 2012
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| Keywords | PE_PGRS62 / Prdx1 / ペルオキシレドキシン / マクロファージ / 結核 |
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| Research Abstract |
PE_PGRS62 is thought to be a virulence factor during tuberculosis. To determine whether PE_PGRS62 is a virulence factor of M. tuberculosis, we deleted PE_PGRS62 of M. tuberculosis Erdman (WT) using mycobacterial phage system. We designed the knockout phage of PE_PGRS62/Rv3812 refer to Erdman genomic DNA. A knockout strain, ΔPE_PGRS62 was constructed and was complimented with the integrative plasmid pMV306 into which PE_PGRS62 gene and its promoter are inserted (Δ62Comp). We also transformed an empty pMV306 as a control strain of Δ62 Comp to ΔPE_PGRS62 (Δ62/mock). To confirm these gene disruption and complementation, we extracted genomic DNA from WT, Δ62/mock and Δ62 Comp strains. The fragments treated with SmaI are detected with specific probe. Next, we examined expression of PE_PGRS62 of WT, Δ62/mock and Δ62Comp strains. These strains were cultured in 7H9/ADC medium and their RNAs were extracted. The transcription levels of PE_PGRS62 were determined by qRT-PCR with specific primers. We observed the recovery of PE_PGRS62 expression in Δ62Comp strain. We checked any difference among WT, Δ62/mock and Δ62Comp. Although we confirmed no differences among these strains grown in 7H9/ADC culture medium, Δ62/mock mutant strain showed reduced the survival rate of intracellular mycobacteria in J774 cells. To evaluate the virulence attenuation of ΔPE_PGRS62 in mice, we injected the strains of WT or ΔPE_PGRS62 into tail vein of mice. There was a significant delay in survival infected with the ΔPE_PGRS62 mutant strain relative to WT strain in BALB/c and SCID mice. These results demonstrate that PE_PGRS62 is required for M. tuberculosis virulence.
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Research Products
(18 results)
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[Journal Article] A novel mechanism of growth phase-dependent tolerance to isoniazid in mycobacteria.2012
Author(s)
Niki M, Niki M, Tateishi Y, Ozeki Y, Kirikae T, Lewin A, Inoue Y, Matsumoto M, Dahl JL, Ogura H, Kobayashi K, Matsumoto S
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Journal Title
J Biol Chem
Volume: 287(33)
Pages: 27743-52
DOI
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[Journal Article] Comprehensive multicenter evaluation of a new line probe assay kit for identification of Mycobacterium species and detection of drug-resistant Mycobacterium tuberculosis.2012
Author(s)
Mitarai S, Kato S, Ogata H, Aono A, Chikamatsu K, Mizuno K, Toyota E, Sejimo A, Suzuki K, Yoshida S, Saito T, Moriya A, Fujita A, Sato S, Matsumoto T, Ano H, Suetake T, Kondo Y, Kirikae T, Mori T
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Journal Title
J Clin Microbiol
Volume: 50(3)
Pages: 884-90
DOI
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