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Expression and Secretion of Serratia marcescens 58 KD Chitinase in Escherichia coli (대장균에서 Serratia marcescens 58KD 키티나아제의 발현과 분비)

  • 장규일;강송옥;신용철
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.511-518
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    • 1992
  • We subcloned a 58 KD chitinase gene of Serratia marcescens into Escherichia coli and investigated the expression and secretion of the chitinase. Chitinase was produced in E. coli by using its own promoter but the levels of enzyme were very low, less than 5 mU/m$\ell$. However, by the combined action of the chitinase and lac promoters, the chitinase activity increased up to about 80 mU/m$\ell$. The most of the chitinase produced in E. coli was localized in periplasm and the small amounts were observed in cytosol and culture medium. Intracellular chitinase activities increased in proportion to the growth of E. coli up to the early stationary phase but rapidly decreased thereafter, which was assumed to be degradation of the chitinase by E. coli proteolytic enzymes.

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Characterization of plasmids of Zymomonas mobils and Construction of E. coli-Zymomonas shuttle Vector (Zymomonas mobilis플라스미드의 특성연구 및 E.coli-Zymomonas셔틀 벡터 제조)

  • 이용억;이병재;강현삼
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.56-63
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    • 1985
  • We have characterized the plasmids of zymomonas, and constructed E. coli-Zymomonas shuttle vector. Plasmids have been detected in four strains of Zymomonas mobilis. All strains tested had at least one plasmid ranging in size from about 1.7 to 46kb. Antibiotics resistances of Z. mobilis were tested to select the host strain. All strains were very sensitive to tetracycline and chloramphenicol. Homology tests between the plasmids in four strains showed that the plasmids of ATCC10988 is highly homologous to those of ZM1, and that there is no homology between plasmids of ZM4 and Agll. The 1.7kb plasmid of ATCC10988, named as pZM886, also has no homology with plasmids of ZM4. A hybrid plasmid, designated to pBZ41, was constructed from pZM886 and pBR322. A restriction map of pBZ41 was established. Replicon of pZM886 didn't operate in E.coli and pBR322 seemed not to replicate in Zymomonas. pBZ41 was transfered from E. coli to Zymomonas by conjugal mobilization. The transconjugants were resistant to tetracycline and maintained pBZ41 stably.

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Antimicrobial Activity of Grapefruit Seed Extract (자몽 종자 추출물의 항균성)

  • Park, Heon-Kuk;Kim, Sang-Bum
    • The Korean Journal of Food And Nutrition
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    • v.19 no.4
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    • pp.526-531
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    • 2006
  • Minimum inhibition concentration(MIC), growth inhibition activity, and colony forming inhibitory activity of grapefruit seed extract against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Escherichia coli, Salmonella enterifidis and Serratia marcescens were tested. MIC of grapefruit seed extract against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Escherichia coli, Salmonella enteritidis and Serratia marcescens was 12.5, 12.5, 12.5, 50, 50, 100ppm, respectively. Growth inhibition concentration of grapefruit seed extract against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Escherichia coli, Salmonella enteritidis and Serratia marcescens was below 1.0, 6.25, below 1.0, 6.25, 25, 25ppm, respectively. Colony forming inhibitory activity of grapefruit seed extract against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Escherichia coli, Salmonella enteritidis and Serratia marcescens was 93.9, 94.0, 99.9, 4.4, 82.7, 86.4%, respectively. Colony forming inhibitory activities of grapefruit seed extract against Gram positive bacteria were higher than that against Gram negative bacteria.

Experimental infection of Campylobacter jejuni and Campylobacter coli in chicks (유추(幼雛)에 있어서 Campylobacter jejuni 및 Campylobacter coli의 실험적(實驗的) 감염(感染))

  • Chung, Byong-gon;Kang, Ho-jo
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.345-353
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    • 1988
  • The present study was carried out to investigate the incidence of C jejuni and C coli in chicken. Also were examined the pathogenicity of the isolates in chick by experimental inoculation. Thermophilic Campylobacter were isolated from 34(45.9%) of the 74 specimens, and classified as 21.6% C jejuni, and 24.3% C coli. In the biotyping of 16 stranis of C jejuni isolates, 37.5% of the strains were grouped as biotype I, 62.5% as biotype II. In the case of 18 strains of C coli isolates, 49.9% of isolates were grouped as biotype I, 55.6% as biotype II. n oral inoculation with $10^4cfu$ of Campylobacter isolates into infant chicks(1 to 3 days-old), 17 days-old and 34 days-old chicks, 32.5% of the chicks developed diarrhea on day 1, 52.5% on day 3, 70.0% on day 5, and 27.5% on day 7, and the peak incidence of diarrhea was reached on day 5. The organisms were found to be discharged in feces one day afterwards. C jejuni and C coli strains were detected from the feces in 87.5% of the chicks on day 5. The organisms were multiplied from $10^4$ to $10^6cfu/gm$ in feces 5 to 7 days after inoculation. C jejuni and C coli recovered from 100% of the cecum, 64.3% of the duodenum, 50.0% of the spleen, 42.9% of the livers, and from 21.4% of gallbladders 7 days after inoculation.

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Comparative study of Clostridium perfringens, Salmonella spp. and E. coli focused on characteristics of E. coli O157 isolated from pigs of HACCP- and non-HACCP-accredited swine farms in Korea

  • Keum, Hyun Ok;Kim, Hye Kwon;Rho, Se Mi;Moon, Hyoung Joon;Park, Seong Jun;Park, Bong Kyun
    • Korean Journal of Veterinary Research
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    • v.50 no.2
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    • pp.113-116
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    • 2010
  • To determine the prevalence of Escherichia (E.) coli O157 : H7 from pigs after the Hazard Analysis and Critical Control Point (HACCP) system has been applied to Korean swine farm since 2006, 291 fecal samples were tested between May and December in 2008. Four E. coli O157:non-H7 (1.4%) were isolated from 4 different non-HACCP-accredited farms and they didn't have virulent genes which can cause illness for human. Also, Clostridium (C.) perfringens, Salmonella spp. and E. coli enterotoxins were tested using multiplex PCR. The positive rate for these pathogens of non-HACCP-accredited farms was higher than that of HACCP-accredited farms, and especially in case of C. perfringens, E. coli enterotoxins LT and STa, it was statistically significant (p < 0.05). Thus, the early implementation of the HACCP program is expected to greatly contribute to the safety of livestock products as well as food hygiene.

Cytokine Gene Expression of Peritoneal Tissues in Response to Mixed Infection of Bacteroides fragilis and Escherichia coli (Bacteroides fragilis와 대장균의 혼합 감염에 의한 복강 조직의 Proinflammatory Cytokine 유전자 발현 조절)

  • Kim, Jung-Mogg;Kim, Young-Jeon;Park, Hwon-Kyum;Cho, Yang-Ja
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.41-48
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    • 2000
  • Bacteroides fragilis and Escherichia coli, normal colonic inhabitants, are the most frequently isolated bacteria in infected tissues, particularly in intraabdominal abscesses. This study was designed to determine whether enteric bacteria may alter the B. fragilis-induced expression of pro inflammatory cytokines in mouse peritoneal tissue (MPT). After C57BL/6 mice were inoculated with abscess-forming mixture containing B. fragilis in the presence or absence of E. coli, RNA was extracted from MPT. Expression of interleukin (IL)-$1{\alpha}$ and tumor necrosis factor $(TNF){\alpha}$ mRNA was assessed using RT-PCR and standard RNA. Each cytokine protein was also measured by ELISA. The co-inoculation of E. coli into mouse peritoneal cavity advanced the onset of abscess development by B. fragilis infection. When mouse was co-infected with E. coli and B. fragilis intraperitoneally, there was a synergistic increase in the expression of IL-$1{\alpha}$ and $TNF{\alpha}$ mRNA in MPT and this was paralleled by increased cytokine protein secretion. Mixed inoculation of heat-killed E. coli and B. fragilis did not cause a synergistic increase in those cytokine mRNA expression. These results suggest that enteric bacteria may significantly affect proinflammatory cytokine signal produced by host peritoneal cavity in response to B. fragilis infection.

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Production of Recombinant Trehalose Synthase from Thermus caldophilus GK24 (재조합 내열성 트레할로스 합성효소의 생산)

  • Choi, Jae-Youl;Cha, Wol-Suk;Shin, Hyun-Jae
    • KSBB Journal
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    • v.21 no.4
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    • pp.298-301
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    • 2006
  • A gene(GeneBank AF 135796) coding for a trehalose synthase from Thermus caldophilus GK24 was cloned into Escherichia coli K12 using five vector systems. The constitutive expression system(pHCETS) which shows the highest trehalose synthase activity from flask culture of recombinant E. coli was selected for the production of trehalose from maltose. For the shake flask culture, the final dry cell weight was 0.9 g/L and the trehalose synthase activity was 25 U/mL. Fed-batch culture of recombinant E. coli harboring plasmid pHCETS which uses the glycerolas a carbon source was performed in jar fermentor: the dry cell weight of 20 g/L and the trehalose synthase activity of 13.7 U/mL were attained in 48 h.

Evaluating the Role of Curcum Powder as a Protective Factor against Bladder Cancer - An Experimental Study

  • El-Mesallamy, Hala;Salman, Tarek M.;Ashmawey, Abeer M.;Osama, Nada
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5287-5290
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    • 2012
  • Throughout human history, plant products have been used for many purposes including as medicines. Herbal products and spices can be used as preventive agents against cancer due to their antimicrobial, antioxidant and antitumorigenic properties. This study was designed to evaluate the potential protective effect of curcum in rats administered nitrosamine precursors; dibutylamine (DBA) and sodium nitrate (NaNO3); and infected with Escherichia coli (E. coli) and also to monitor changes in nuclear factor the Kappa B p65 (NF-${\kappa}B$ p56) pathway and its downstream products, Bcl-2 and interleukin-6 (IL-6), in parallel with nitrosamine precursors, E. coli and curcum treatment. Rats were divided into three groups (n=25 each; except of control group, n+20). Group I a normal control group, group II administered DBA/NaNO3 in drinking water and infected with E. coli and group III was administered DBA/NaNO3 in drinking water, infected with E. coli and receiving standard diet containing 1% curcum powder. Histopathological examination reflected that the curcum treated group featured a lower incidence of urinary bladder lesions, and lower levels of NF-${\kappa}B$, Bcl-2 and IL-6, than the group receiving nitrosamine precursor and infected with E. coli. These findings suggested that curcum may have a protective role during the process of bladder carcinogenesis by inhibiting the NF-${\kappa}B$ pathway and its downstream products.

Antimicrobial Resistance of Escherichia coli Isolates from Mussel Mytilus galloprovincialis Farms and Inland Pollution Sources in the Changseon Area, Korea (창선 해역의 지중해담치(Mytilus galloprovincialis) 양식장 및 육상오염원에서 분리한 대장균(Escherichia coli)의 항생제 내성)

  • Kwon, Soon Jae;Lee, Ka Jeong;Jung, Yeoun Joong;Park, Sang Gi;Go, Kyeong Ri;Yang, Ji Hye;Mok, Jong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.564-572
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    • 2016
  • In this study, we isolated and characterized Escherichia coli from mussels and inland pollution sources in or in proximity to the Changseon area on the southern coast of Korea in 2014. A total of 147 strains of E. coli were isolated from 54 mussels and 32 pollution-source samples. The susceptibility of the isolates to 24 antimicrobial agents was analyzed. The resistance of E. coli isolates to rifampin was highest at 100%, followed by cephalothin (98.6%), tetracycline (91.8%), amikacin (81.0%), ampicillin (79.6%), cefazolin (79.6%), streptomycin (73.5%), piperacillin (70.7%), gentamicin (37.4%), cefoxitin (35.4%), cefamandole (34.7%), tobramycin (29.9%), amoxicillin/clavulanic acid (24.5%), nalidixic acid (21.8%), trimethoprim (19.0%), chloramphenicol (17.7%), cefotaxime (12.9%), trimethoprim (10.9%), ceftazidime (10.2%), aztreonam (7.5%), imipenem (2.7%), cefepime (2.0%), and cefotetan (0.0%). In addition, the antimicrobial resistance of E. coli isolates from inland pollution sources was slightly greater than or similar to that of isolates from mussels.

Antimicrobial Resistance Patterns of Escherichia coli and Vibrio parahaemolyticus Isolated from Shellfish from the West Coast of Korea (서해안 패류에서 분리한 대장균 및 장염비브리오균의 항균제 내성 패턴)

  • Ryu, A Ra;Park, Kunbawui;Kim, Song Hee;Ham, In Tae;Kwon, Ji Young;Kim, Ji Hoe;Yu, Hong Sik;Lee, Hee Jung;Mok, Jong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.6
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    • pp.662-668
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    • 2017
  • This study investigated the antimicrobial resistance patterns of Escherichia coli and Vibrio parahaemolyticus isolated from oysters Crassostrea gigas, short-necked clams Ruditapes philippinarum and corb shells Cyclina sinensis from the West Coast of Korea from June through November 2013. The antimicrobial susceptibility patterns of the isolated strains of E. coli and V. parahaemolyticus to 12 antimicrobial agents used in Korea for clinical or veterinary therapy were analyzed. Antimicrobial resistance to at least one antibiotic was seen in 52.0% of the E. coli isolates (156 strains) and 44.3% of the V. parahaemolyticus isolates (194 strains). The resistance of the E. coli (34.0%) and V. parahaemolyticus (41.8%) isolates to ampicillin was highest. Multiple antimicrobial resistance against at least three antimicrobials was seen in 9.0% of the E. coli isolates and 1.0% of the V. parahaemolyticus isolates.