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Neisseria gonorrhoeae PID332

ABSTRACT: Clinical isolate

PROVIDER: PRJNA32803 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR004141.fastq.gz	Fastqsanger.gz
	SRR032789.fastq.gz	Fastqsanger.gz
	SRR136774.fastq.gz	Fastqsanger.gz

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Neisseria gonorrhoeae 1291

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Project description:Neisseria gonorrhoeae (NG) exhibits high genome plasticity caused by an unusually high density and diversity of transposable elements, and easily performs various mechanisms of drug resistance. Here we investigated the i19.05 clinical isolate with reduced susceptibility to penicillin (MIC=0.5 mg/L), tetracycline (MIC=0.5 mg/L), and azithromycin (MIC=1.0 mg/L), which carried no known genetic resistance determinants except of penA, which cannot explain the expression of the resistant phenotype. In addition, it attracted our attention to the presence of a new and unique mutation of Asn105Ser in SurA and several mutations in Omp85 (BamA). The goal of our study was to search for new molecular mechanisms of drug resistance. The pan susceptible n01.08 NG clinical isolate was involved as a control to compare, as well as a recipient in transformation procedure. The fragments of i19.05 genome contained mutant surA, omp85, and penA genes were amplified and used in spot-transformation of the n01.08 recipient isolate as described (Ilina, 2013). Finally, a resistant transformant NG05 (PenAmut, Ompmut, SurAmut) was obtained. For comprehensive proteomic analysis via LC-MS/MS, the proteins from the all tested N. gonorrhoeae strains were fractionated on cell envelope (CE) (including outer membrane, periplasmic, inner membrane) and cytosol (C). A total of 1125 proteins in the CE fraction, of which 894 were common in all strains were identified. Proteomics of the C fraction in the same experiment yielded a total of 928 proteins, of which 676 were shared among all strains. Proteome coverage for both fractions ranged from 52.72% (1111 proteins) in n01.08 to 54.53% (1149 proteins) in i19.05.

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Neisseria gonorrhoeae

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Neisseria gonorrhoeae drg::cm versus wild type and Neisseria gonorrhoeae drg::dam versus wild type

Project description:Transcriptional profiling of N. gonorrhoeae comparing wild type cells to cells with inactivated by chloramphenicol cassette (cm) dam replacing gene (drg) or wild type cells comparing to cells with inserted dam gene. The Goal was to study the role of drg or dam presence in overall expression profile.

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Project description:Neisseria gonorrhoeae, the etiologic agent of gonorrhea, is frequently asymptomatic in women, often leading to chronic infections. One factor contributing to this may be biofilm formation. N. gonorrhoeae can form biofilms over glass and plastic surfaces. There is also evidence that biofilm formation may occur during natural cervical infection. To further study the mechanism of this biofilm formation, transcriptional profiles of N. gonorrhoeae biofilm were compared to planktonic profiles. Biofilm RNA was extracted from N. gonorrhoeae 1291 grown for 48 hours in continuous flow chambers over glass. Planktonic RNA was extracted from the biofilm runoff. When biofilm was compared to planktonic growth, 3.8 % of the genome was differentially regulated. Genes highly up-regulated in biofilm included aniA, norB, and ccp, which play critical roles in anaerobic metabolism and oxidative stress tolerance. Down-regulated genes included the nuo gene cluster (NADH dehydrogenase) and the cytochrome bcI complex, which are involved in aerobic respiration and are thought to contribute to endogenous oxidative stress. Furthermore, we determined that aniA, ccp, and norB insertional mutants are attenuated for biofilm formation over glass and transformed human cervical epithelial cells (THCEC). This data suggests that biofilm formation could minimize oxidative stress during cervical infection and allow N. gonorrhoeae to maintain a nitric oxide steady state that may be anti-inflammatory.

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