• Title/Summary/Keyword: MudJ operon fusion

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Osmotic-inducible Gene Expression using MudJ(Km.lac) Operon Fusion in Salmonella typhimurium (Salmonella typhimurium에서 MudJ(Km.lac) 오페론 융합을 이용한 삼투유도유전자의 발현)

  • 주성관;우영대;허연주;안정선;박용근
    • Korean Journal of Microbiology
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    • v.29 no.4
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    • pp.215-220
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    • 1991
  • MudJ(Km.lac) operon fusions were used in the identification of osmotic-inducible genetic(osi) loci in Salmonella typhimurium. Expression of osi::lacZ(osi5001, 5027) genes were dramatically induced 39-189 fold when the osmolarity was increased. Seven osm::lacZ genes were constituvely expressed under both low and high osmotic strength. The osi5001::lacZ fusion strains showed the enhanced osmotolerance and the reduced expression of the osi5001::lacZ in the presence of 1mM proline or betaine as osmoprotectants. Four osmotic inducible genetic loci were mapped into 36 (YK531), 44 (YK504), 57 (YK501) and 84 (YK528) map unit by testing the cotransduction frequency.

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Genetic Responses to Metal ion in Aslmonella typhimurium (Salmonella typhimurium의 금속이온에 대한 유전적 반응)

  • Jung, Ju-Ri;Park, Kyeong-Ryang;Koh, Sang-Kyun;Park, Yong-Keun;Lee, In-Soo
    • Journal of Life Science
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    • v.8 no.2
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    • pp.216-225
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    • 1998
  • Metal ion-induced and it’s regulatory genes were screened in virulent salmonella typhimurium UK1 and tested cross-regulation with various stresses. Using the techniqud of P22-MudJ(Km, lacZ)-directed lacZ operon fusion, LF40 cuiA::MudJ and Lf153 cuiD::MudJ which were induced by copper were selected. cuia and cuiD were determined anaerobic coper inducible and copper tolerance response gene, respectively. Also cuiA and cuiD locus were determined at 81 and 8min, respectively, on salmonella Genetic Map. The two regulators were identified as cuaR, and cudR, which controls cuiA and cuiD, respectively. cuaR, and cudR appeared as negative regulators because the expression of cuiA-lac-Z and cuiD-lacZ were increased. Copper adapted UK1 showed high resistance to H$_{2}$O$_{2}$, but cuiD did not. The product of the cudR locus was responsible for decreasing the tolerance to copper and H$_{2}$O$_{2}$. Furthemore cuiA and cuiD locus were found to be part of a regulon under the control of a trans-acting regulators, rpoS, oxyR and relA. Therefore, the results suggest CTR participate with oxidative stress on Salmonella.

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Anaerobic Acid Tolerance Response in Salmonella typhimurium (Salmonella typhimurium의 혐기적 산내성도 평가)

  • Kim, Young-Chan;Lee, Sun;Lee, Kyung-Mi;Im, Sung-Young;Park, Yong-Geun;Baek, Hyung-Seok;Park, Kyung-Ryang;Lee, In-Soo
    • Journal of Life Science
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    • v.9 no.2
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    • pp.169-175
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    • 1999
  • Salmonella typhimurium can encounter a wide variety of environments during its life cycle. In nature, S. typhimurium can experience and survive dramatic acid stresses that occur in diverse ecological niches ranging from pond water to phagolysosomes. These survival mechanism is aquired by the Acid Tolerance Response(ATR) in Salmonella. The ATR of S. typhimurium is a complex inducible phenomenon in which exposures to slight or moderate low pH will produce a stress response capable of protecting the organism against more severe acid challenges. ATR in Salmonella has two different systems that are called RpoS dependent and independent. We found that ATR in anaerobic was showed RpoS independent because rpoS$\Omega$AP had ATR as S. typhimurium UK1. Using the P22 MudJ(Km, lacZ) operon fusion technique and a lethal selection procedure combining low pH(pH4.5) and sodium acetate(10mM, pH4.5), we isolated LF487 aatA::MudJ which showed acid sensitive in anaerobic condition. aatA locus was determined at 12 min on Salmonella Genetic Map. The survival rate of aatA mutant was showed significantly diminished at pH4.3 than virulent wild type Salmonella in anaerobic condition(5% $CO_2$, 5% H$_2$, 90% $N_2$). Therefore isolated gene was confirmed important gene for anaerobic ATR system.

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Low Temperature Inducible Acid Tolerance Response in virulent Salmonella enterica serovar Typhimurium (병원성 Salmonella enterica serovar Typhimurium의 저온 유도성 산 내성 반응)

  • Song, Sang-Sun;Lee, Sun;Lee, Mi-Kyoung;Lim, Sung-Young;Cho, Min-Ho;Park, Young-Keun;Park, Kyeong-Ryang;Lee, In-Soo
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.228-233
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    • 2001
  • The acid tolerance response (ATR) of log-phase Salmouella enterica seroyar Typhimurium is induced by acid adaptation below pH4.5 and will protect cells against more severe acid. Two distinctive ATR systems in thisorganism are a log-phase and stationary-phase ATR in which acid adaptations trigger the synthesis of acid shockproteins (ASPs). We found that log-phase ATR system was strongly affected by environmental factor, low tem-perature, $25^{\circ}C$. Exposure to low temperature and mild acid has been shown to increase acid survival dra-matically, and this survival rate was showed higher than $37^{\circ}C$. Especially unadapted cells at $25^{\circ}C$ presented tenthousand folds survival increasing when compared with cells at $37^{\circ}C$. The degree of acid tolerance of rpoSwhich is blown to be required for acid tolerance more increase than $37^{\circ}C$. Even though AIR pattern of rpoSbetween unadapted and adapted was showed similar at pH 3.1, rpoS-dependent ATR system also has beendetected in low temperature because rpoSAp prevents sustained acid survival at $25^{\circ}C$. Therefore the resultssuggest low temperature ATR system requires rpoS-dependent and -independent both. To investigate the basisfor low temperature related ATR system, gene that was participated for low temperature acid tolerance (lat) wasscreened in virulent S. enterica serovar Typhimurium UKl Using the technique of P22- MudJ (Km, lacZ)-directed lacZ operon fusion, LF452 latA‥‥MudJ was isolated. The latA‥‥MudJ of S. enterica Typhimurium pre-vented low temperature acid tolerance response. Therefore latA is considered one of the important genes for acidadaptation.

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