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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제25권 제10호
- 발행연도
- 2015.1
- 수록면
- 1,621 - 1,628 (8page)
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초록· 키워드
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Chlamydia possesses a conserved 7.5 kb plasmid that is known to play an important role in chlamydial pathogenesis, since some chlamydial organisms lacking the plasmid are attenuated. The chlamydial transformation system developed recently required the use of plasmid-free organisms. Thus, the generation and identification of plasmid-free organisms represent a key step in understanding chlamydial pathogenic mechanisms. A tricolor immunofluorescence assay for simultaneously detecting the plasmid-encoded Pgp3 and whole organisms plus DNA staining was used to screen C. muridarum organisms selected with novobiocin. PCR was used to detect the plasmid genes. Next-generation sequencing was then used to sequence the genomes of plasmid-free C. muridarum candidates and the parental C. muridarum Nigg strain. We generated five independent clones of plasmid-free C. muridarum organisms by using a combination of novobiocin treatment and screening plaque-purified clones with anti-Pgp3 antibody. The clones were confirmed to lack plasmid genes by PCR analysis. No GlgA protein or glycogen accumulation was detected in cells infected with the plasmid-free clones. More importantly, whole-genome sequencing characterization of the plasmid-free C. muridarum organism and the parental C. muridarum Nigg strain revealed no additional mutations other than loss of the plasmid in the plasmid-free C. muridarum organism. Thus, the Pgp3-based immunofluorescence assay has allowed us to identify authentic plasmid-free organisms that are useful for further investigating chlamydial pathogenic mechanisms.
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참고문헌 (22)
참고문헌 신청
Borges V / 2013 / Effect of long-term laboratory propagation on Chlamydia trachomatis genome dynamics / Infect. Genet. Evol 17 : 23 ~ 32
Carlson JH / 2008 / The Chlamydia trachomatis plasmid is a transcriptional regulator of chromosomal genes and a virulence factor / Infect. Immun 76 : 2273 ~ 2283
Chen C / 2006 / The hypothetical protein CT813 is localized in the Chlamydia trachomatis inclusion membrane and is immunogenic in women urogenitally infected with C. trachoma
Chumduri C / 2013 / Chlamydia infection promotes host DNA damage and proliferation but impairs the DNA damage response / Cell Host Microbe 13 : 746 ~ 758
Frazer LC / 2012 / Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection / PLoS. One 7 :
Carlson JH / 2008 / The Chlamydia trachomatis plasmid is a transcriptional regulator of chromosomal genes and a virulence factor / Infect. Immun 76 : 2273 ~ 2283
Chen C / 2006 / The hypothetical protein CT813 is localized in the Chlamydia trachomatis inclusion membrane and is immunogenic in women urogenitally infected with C. trachoma
Chumduri C / 2013 / Chlamydia infection promotes host DNA damage and proliferation but impairs the DNA damage response / Cell Host Microbe 13 : 746 ~ 758
Frazer LC / 2012 / Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection / PLoS. One 7 :
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