• Title/Summary/Keyword: tetK

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Functional Expression of SAV3818, a Putative TetR-Family Transcriptional Regulatory Gene from Streptomyces avermitilis, Stimulates Antibiotic Production in Streptomyces Species

  • Duong, Cae Thi Phung;Lee, Han-Na;Choi, Si-Sun;Lee, Sang-Yup;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.136-139
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    • 2009
  • Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-and-constitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the low-producer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species.

Optimal Food and Concentration for the Growth of the Ultra-small Rotifer Synchaeta kitina (초소형 Rotifer Synchaeta kitina의 성장을 위한 최적 먹이 및 공급량)

  • Park, Jin-Chul;Park, Heum-Gi
    • Journal of Aquaculture
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    • v.21 no.2
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    • pp.76-81
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    • 2008
  • We investigated the food-effect for ultra-small rotifer Synchaeta kitina cultured under a individual and community by several diets: 3 single trials(Tetraselmis suecica, TET; Isochrysis galbana, ISO; Marine Chlorella ellipsoidea, CHL) and 3 trials with a mixture of 2 species. The rotifer was cultured on the different feeding concentrations. In the individual cultures, the maximum number of offsprings and maximum lifespan of the female investigated to 5.8 inds. and 12.7 days in TET trial, respectively. Values of the developmental phases of the rotifer fed with T. suecica were higher than those of trials without T. suecica. Also it approached faster to maturation level. In the community cultures, the maximum density of TET+CHL trial elevated up to 1,569 inds./mL. But, CHL and ISO showed a poor growth rate and maximum density. The offsprings of the female increased continuously when fed by T. suecica trial, up to $10{\times}10^3$ cells/ind./day. As the quantity of supplied diet was lowered their lifespan were decreased. But, the maximum density and growth rate in the community cultures showed the highest value in the $10{\times}10^3$ cells/ind./day. The efficient food for mass culture of S. kitina was T. suecica, and optimum concentration of their food was 10,000 cells for an individual.

PERIODONTOPATHIC BACTERIA AND ANTIBIOTIC RESISTANCE GENES OF ORAL BIOFILMS IN CHILDREN (어린이 치면세균막에서 치주질환원인균과 항생제 내성유전자의 출현율)

  • Kim, Seon-Mi;Choi, Nam-Ki;Cho, Seong-Hoon;Lee, Seok-Woo;Lim, Hoi-Jeong;Lim, Hoi-Soon;Kang, Mi-Sun;Oh, Jong-Suk
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.2
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    • pp.170-178
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    • 2011
  • The purpose of this study was to assess the prevalence of periodontopathic bacteria and resistance determinants from oral biofilm of children. Subgingival dental plaque was isolated from 87 healthy children, and PCR was performed to determine the presence of 5 periodontal pathogens including P. gingivalis, T. forsythia, T. denticola, F. nucleatum, A. actinomycetemcomitans, and nine resistance genes including tet(Q), tet(M), ermF, aacA-aphD, cfxA, $bla_{SHV}$, $bla_{TEM}$, vanA, mecA. 1. The prevalence of F. nucleatum, T. forsythia. and P. gingivalis was 95.4%, 55.2%, and 40.2%, respectively. In addition. the prevalence of A. actinomycetemc omitans was 5.7%, while T. denticola was 3.4%. 2. In analysis of antibiotic resistance determinants. cfxA, $bla_{TEM}$ and tet(M) were detected in all the samples tested. It was also found that the prevalence of tet(Q) showing tetracycline resistance. $bla_{SHV}$ associated with resistance to ${\beta}$-lactams, ermF exhibiting erythromycin resistance, and, vanA resulting vancomycin resistance was 88.5%, 29.9% 87.4%, and 48.5%, respectively. The aacA-aphD gene showing resistance to aminoglycosides and mecA gene harboring methicillin resistance exhibited the lowest prevalence with 9.2%. 3. In a correlation analysis between periodontopathic pathogens and antibiotic resistance determinants, it was found that there was a significant correlation between T. forsythia and $bla_{SHV}$. Also, P. gingivalis and vanA showed a correlation. Finally, tet(Q) and ermF showed a significant correlation (phi: 0.514) while mecA and vanA also showed a correlation(phi: 0.25).

Veterinary antibiotic oxytetracycline's effect on the soil microbial community

  • Danilova, Natalia;Galitskaya, Polina;Selivanovskaya, Svetlana
    • Journal of Ecology and Environment
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    • v.44 no.2
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    • pp.72-80
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    • 2020
  • Background: Antibiotics are widely used to treat animals from infections. After fertilizing, antibacterials can remain in the soil while adversely affecting the soil microorganisms. The concentration of oxytetracycline (OTC) in the soil and its effect on the soil microbial community was assessed. To assess the impact of OTC on the soil microbial community, it was added to the soil at concentrations of 50, 150, and 300 mg kg-1 and incubated for 35 days. Results: The concentration of OTC added to the soil decreased from 150 to 7.6 mg kg-1 during 30 days of incubation, as revealed by LC-MS. The deviations from the control values in the level of substrate-induced respiration on the 5th day of the experiment were, on average, 26, 68, and 90%, with OTC concentrations at 50, 150, and 300 mg kg-1, respectively. In samples with 150 and 300 mg kg-1 of OTC, the number of bacteria from the 3rd to 14th day was 2-3 orders of magnitude lower than in the control. The addition of OTC did not affect the fungal counts in samples except on the 7th and 14th days for the 150 and 300 mg kg-1 contaminated samples. Genes tet(M) and tet(X) were found in samples containing 50, 150, and 300 mg kg-1 OTC, with no significant differences in the number of copies of tet(M) and tet(X) genes from the OTC concentration. Conclusions: Our results showed that even after a decrease in antibiotic availability, its influence on the soil microbial community remains.

Induction of Sex Maturation and Growth in Comb Pen Shells, Atrina pectinata per Microalgae Classes (미세조류 종류에 따른 키조개, Atrina pectinata의 성장 및 성숙 유도)

  • Moon, Tae-Seok;Jo, Pil-Gue;Kim, Byoung-Hak;Park, Ki-Yeol;Ku, Hag-Dong;Shin, Yun-Kyung;Lym, Young-Sub
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.105-112
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    • 2009
  • We investigated the degree of obesity, histological development stages of gonads and sexual maturation induction rates of comb pen shell, Atrina pectinata, per the type of micro-algae supplied. Terms of maturation by singular or mixed supply of microalgae, it was found that maturation of the female was the quickest at 60.0% by the Tetraselmis tetrathele (Tet). experiment group followed by 57.1% by the Chlorella ellipsoidea (Chl). experiment group and 16.7% by the Phaeodactylum tricornutum (Pha). experiment group. However, there were no significant differences between Tet. experiment group and Chl. experiment group. As for the male, maturation was the quickest at 60.0% by the Tet. experiment group followed by 16.7% by the Chl. experiment group and 14.3% by the Pha. experiment group. In light of these results, Tet. is concluded to be a very useful feed organism in breeding the mother comb pen shells. Upon completion of the experiment, the sexual maturation induction rate for the female was found to be the highest at 82.0% in the Tet. experiment group followed by 72.0% by the Chl. experiment group, 64.0% by the Pha. experiment group and 58.0% by the mixed micro-algae experiment group. During the period of experiment, the survival rate was the highest at 94.4% by the mixed micro-algae experiment group followed by 90.0% by the Pha. experiment group, 83.1% by the Tet. experiment group and 78.8% by the Chl. experiment group.

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Induction of cancer cell-specific death via MMP2 promoterdependent Bax expression

  • Seo, Eun-Jeong;Kim, Se-Woon;Jho, Eek-hoon
    • BMB Reports
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    • v.42 no.4
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    • pp.217-222
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    • 2009
  • Controlled gene expression in specific cells is a valuable tool for gene therapy. We attempted to determine whether the lentivirus-mediated Tet-On inducible system could be applied to cancer gene therapy. In order to select the genes that induce cancer cell death, we compared the ability of the known pro-apoptotreic genes, Bax and tBid, and a cell cycle inhibitor, p21cip1/waf1, and determined that Bax was the most effective. For the cancer cell-specific expression of $rtTA2^S$-M2, we tested the matrix metalloproteinase-2 (MMP-2) promoter and determined that it is highly expressed in cancer cell lines, including SNU475 cells. The co-transduction of two lentiviruses that contain sequences for TRE-Bax and $rtTA2^S$-M2, the expression of which is controlled by the MMP-2 promoter, resulted in the specific cell death of SNU475, whereas other cells with low MMP-2 expression did not evidence significant cell death. Our data indicate that the lentivirus-mediated Tet-On system using the cancer-specific promoter is applicable for cancer gene therapy.

The Effects of Peroxiredoxin III on Human HeLa Cell Proliferation

  • Choi, Soonyoung;Kang, Sangwon
    • IMMUNE NETWORK
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    • v.3 no.4
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    • pp.276-280
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    • 2003
  • Background: Peroxidases (Prx) of the peroxiredoxin family reduce hydrogen peroxide and alkyl hydroperoxides to water and alcohol respectively. Hydrogen peroxide is implicated as an intracellular messenger in various cellular responses such as proliferation and differentiation. And Prx I activity is regulated by Cdc-2 mediated phosphorylation. This work was undertaken to investigate the proliferation role of peroxiredoxin III as a member of Prx family in Prx III overexpressed HeLa cell line. Methods: To provide further evidence of proliferation, we selected Prx III stably expressed HeLa Tet-off cell lines. Cell proliferation was examined by using proliferation reagent WST-1 in the presence or absence of doxycycline. Prx III, 2-cys Prx enzymes exist as homodimer. The activation of Prx III heterodimer with induced and endogenous Prx III was examined by immunoprecipitation. Results: Immunoprecipitation analysis of the induced and endogenous Prx III with anti-myc showed that the induced wild type (WT) and dominant negative (DN) Prx III from HeLa Prx III Tet-off stable cell heterodimerized with endogenous Prx III each other. And the expression level of induced Prx III was examined after addition of doxycycline. By 72 hr, the expression level of induced Prx III was diminished gradually and the half-life of the induced wild type Prx III was approximately 17 hr. The proliferation experiment demonstrated that the relative proliferation value of induced and endogenous WT Prx III stable cell has no changes but the DN Prx III induced HeLa Tet-off stable cells were lower than endogenous Prx III. Conclusion: In conclusion, the HeLa dominant negative Prx III Tet-off stable cells were decreased the proliferation.

Antimicrobial Resistance and Virulence Genes Presence in Escherichia coli Strains Isolated from Gomso Bay, Korea

  • Park, Kwon-Sam
    • Fisheries and Aquatic Sciences
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    • v.16 no.4
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    • pp.221-227
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    • 2013
  • In total, 131 Escherichia coli isolates from surface seawater of the Gomso Bay, of Korea, were analyzed for their susceptibility to 22 different antimicrobials and for genes associated with antimicrobial resistance and virulence. According to the disk diffusion susceptibility test, the resistance to tetracycline was most prevalent (33.6%), followed by that to ampicillin (22.1%), ticarcillin (22.1%), and trimethoprim (16.8%). More than 46.6% of the isolates were resistant to at least one antimicrobial, and 22.9% were resistant to three or more classes of antimicrobials; these were consequently defined as multidrug resistant. We further found that 29 ampicillin-resistant isolates possessed genes encoding TEM-type (93.1%) and SHV-type (6.9%) ${\beta}$-lactamases. Among the 44 tetracycline-resistant isolates, tetA and tetC were found in 35 (79.5%) and 19 (43.2%), respectively, whereas tetB was detected in only three isolates (6.8%). With regard to virulence genes, merely 0.8% (n = 1) and 2.3% (n = 3) of the isolates were positive for the enteroaggregative E. coli-associated plasmid (pCVD432) gene and the enteropathogenic E. coli-specific attaching and effacing (eae) gene, respectively. Overall, these results not only provide novel insight into the necessity for seawater sanitation in Gomso Bay, but they help reduce the risk of contamination of antimicrobial-resistant bacteria.

Serotype and antimicrobial susceptibility of Actinobacillus pleuropneumoniae isolates from pigs in Korea (돼지에서 분리한 Actinobacillus pleuropneumoniae의 혈청형 분포 및 항생제 감수성)

  • Jung, Ji-Youl;Jang, Hyun
    • Korean Journal of Veterinary Research
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    • v.52 no.3
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    • pp.177-181
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    • 2012
  • Actinobacillus (A.) pleuropneumoniae is the causative agent of pleuropneumonia which is one of the most important respiratory diseases in pigs worldwide. A total of 32 A. pleuropneumoniae isolates from diseased pigs during 2008 to 2010 were serotyped by polymerase chain reaction method. The susceptibility of the isolates to 13 antimicrobial agents were determined by disk diffusion test. In all the 32 isolates examined in this study, serotype 5 (16 isolates: 50%), 1 (7 isolates: 21.9%), 2 (5 isolates: 15.6%) and 12 (1 isolate: 3.1%) were found. Of all tested antimicrobial agents, resistance to oxytetracycline was found in 96.9% of isolates, followed by resistance to amikacin (81.2%), neomycin (68.7%), kanamycin (53.1%), penicillin (50.0%), gentamicin (43.7%), florfenicol (25.0%), ampicillin (18.7%), colistin (9.4%), trimethoprim/sulfamethoxazole, ceftiofur (8.3%), amoxicillin/clavulanic acid (3.1%) and enrofloxacin (0%). Oxytetracycline or florfenicol-resistant isolates were examined for the presence of resistance gene. Among the 31 oxytetracycline-resistant isolates, tetB, tetH and tetO genes were detected in 22 (71%), 8 (26%) and 1 (3%) isolates, respectively. The floR genes were detected in 8 (100%) of the 8 florfenicol-resistant A. pleuropneumoniae isolates.