• Title/Summary/Keyword: operon

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Biotechnology for the Production of Threonine Production (Threonine의 생물공학적 생산)

  • Kim, Kyoung-Ja
    • YAKHAK HOEJI
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    • v.34 no.6
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    • pp.447-456
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    • 1990
  • Various methods are available for the production of L-threonine. The microbial production of L-threonine has been achieved by breeding L-threonine analog-resistant auxotrophic mutants of various bacteria. The enzymatic production of L-threonine has been demonstrated by use of threonine metabolic enzymes such as threonine deaminase, threonine aldolase, or threonine dehydrogenase complex. Threonine synthesis from glycine and ethanol seems to be catalyzed by the enzymes Methanol dehydrogenase(MDH) and Serine hydroxymethyltransferase(SHMT), which was also found to catalyze the aldol condensation of glycine with acetaldehyde. The improved production of L-threonine has been achieved by amplifying the genes for the L-threonine biosynthetic enzymes using recombinant DNA techniques.

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Fnr, NarL and NarP Regulation and Time Course Expression of Escherichia coli aeg-46.5 Gene

  • Ahn, Ju-Hyuk;Choe, Mu-Hyeon
    • BMB Reports
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    • v.29 no.1
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    • pp.88-91
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    • 1996
  • The anaerobically expressed gene aeg-46.5, which had been identified by the operon fusion technique with a hybrid bacteriophage of ${\lambda}$ and Mu, ${\lambda}$placMu53, was studied for its expression pattern and growth. The expression of aeg-46.5 was studied in the wild-type cell and mutant cells that have mutation (s) in the control gene of anaerobic respiration (fnr) and nitrate response (narL and narP). The ${\beta}$-galactosidase reporter gene showed maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Both 40 mM and 100 mM concentrations of nitrate ion in the medium had little effect on expression level. We propose that aeg-46.5 is subject to multiple regulations of anaerobic activation by Fnr, nitrate activation by NarP and repression mediated by NarL.

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Transcriptional Regulatory System by CRP (CRP의 전사 조절계)

  • 최용락
    • Journal of Life Science
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    • v.2 no.1
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    • pp.2-10
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    • 1992
  • cAMP-CRP complex에 의해 전사조절을 받는 관련유전자의 구조, 특징, 조절양상, 기작과 CRP의 생화학적 및 분자생물학적이해 등을 살폈다. 그러나 대장균의 전체유전자의 구조해석이 연구사업으로 실행되어지고 있는 현실에서 볼 때 CRP의 조절하에 있는 유전자군이 보다 더 복잡 다양한 system으로 생각되어져서 CRP조절계의 전체적인 양상으로 해명하기 위한 연구도 이루어질 것으로 생각되어진다. 즉, cAMP-CRP의 조절하에 있는 조절영역의 전체를 파악함은 operon의 발현조절의 해명에 중요한 기틀이 될 뿐만 아니라 개별유전자의 이해를 위한 것보다 복잡한 세포 전체의 제어계를 분자적수준에서 해결하는데 중요한 과제라고 사료되어진다. 몇 가지 연구 결과에서는 cAMP-CRP가 결합은 하면서도 발현조절에는 관여하지 않는다든지, cAMP가 결합하지 않으면서도 전사조절을 받을 수 있는 유전자들이 밝혀지고 있다.

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Mutagenicity and Hepato-Toxicity of Kyoaesamultang (교애사물탕의 변이원성 및 간독성에 관한 연구)

  • 우덕안;홍희탁;문진영;이태균;김철호;김준기;최미정;남경수
    • Toxicological Research
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    • v.13 no.3
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    • pp.197-202
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    • 1997
  • Kyoaesamultang(KAT) has been used as an important prescription for various diseases including threatened abortion, associated with pregnancy in traditional medicine. In oder to identify the safety of KAT, this study was designed to determine mutagenicity and hepato-toxicity. In Rec-assay, Bacillus subtills H-17($Rec{^+}$) and M-45($Rec{^-}$) strains were used to clarify the DNA damage property. In Ames test, Salmonella typhimurium TA98 and TA100 were used for mutagenicity testing. In SOS umu test, Salmonella typhimurium TA1535 containing plasmid pSK1002 was used as a tester strain, and the levels of umu operon expression were monitored by measuring the $\beta$-galactosidase activity. From tested results, KAT did not show DNA damage and mutagenicity. On the other hand, hepato-toxicity of KAT to female ICR mice was monitored by the measurements of s-GOT, s-GPT and LDH activities after oral feeding for 15days. KAT showed 34% increase of s-GOT and s-GPT activities, also exhibited 35% increase of LDH activity in mice sera.

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Application of RAPD Methods in Meat for Beef Breed Identification

  • Choy, Y.H.;Oh, S.J.;Kang, J.O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1655-1658
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    • 2001
  • Bovine genome samples were collected from meat of three different beef breeds (Hanwoo, Holstein and imported beef breed) that are commercially merchandized in Korean beef market. Operon B (OPB)-kits were used as random primers (3, 7, 10, 11, 12, 14) in random amplified polymorphic DNA (RAPD) method on whole genome. Each primer provided characteristic bands that were highly polymorphic. Each single primer could provide relatively efficient polymorphic band patterns among breeds. However, use of two or more primers in combination is recommended to improve resolution of experiments with higher molecular weight bands of DNA. In our experiments, OPB-11 resolved well between beef cattle breeds and Holstein. And OPB-7, 12 and 14 could be combined with OPB-11 to identify Hanwoo beef from the other two kinds of beef.

Regulation of fpr Gene Encoding NADPH : Ferredoxin Oxidoreductase by the soxRS Locus in Escherichia coli

  • Koh, Young-Sang;Choih, Jenny;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.137-143
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    • 1996
  • We isolated a promoter inducible by paraquat, a superoxide-generating agent, from Escherichia coli using a promoter-probing plasmid pRS415. From sequence analysis we found out the promoter is for fpr ENCODING nadph : ferredoxin oxidoreductase. We constructed on operon fusion of lacZ gene with fpr promoter to monitor the expression of the gene in the single-copy state. LacZ expression generators, menadione and plumbagin, also induced the expression of .betha.-galactosidase in the fusion strain. On the other hand, no significant induction was observed by treatment with hydrogen peroxide, ethanol, and heat shock. Induction of .betha.-galactosidase was significantly reduced by introducing a .DELTA. sox 8 :: cat of soxS3 :: Tn10 mutation into the fusion strain, indicating that fpr gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis. Possible roles of fpr induction in superoxide stress were discussed.

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Proteolysis of the Reverse Transcriptase of Hepatitis B Virus by Lon Protease in E. coli

  • Han, Joo-Seok;Park, Jae-Yong;Hwang, Deog-Su
    • Animal cells and systems
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    • v.5 no.3
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    • pp.195-198
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    • 2001
  • Hepatitis B virus (HBV) polymerase, which possesses the activities of terminal binding, DNA polymerase, reverse transcriptase and RNaseH, has been shown to accomplish viral DNA replication through a pregenomic intermediate. Because the HBV polymerase has not been purified, the expression of HBV polymerase was examined in an E. coli expression system that is under the regulation of arabinose operon. The expressed individual domain containing terminal binding protein, polymerase, or RNaseH turned out to be insoluble. The activities of those domains were not able to be recovered by denaturation and renaturation using urea or guanidine-HCI. The expressed reverse transcriptase containing the polymerase and RNaseH domains became extensively degraded, whereas the proteolysis was reduced in a Ion- mutant. These results indicate that Lon protease proteolyzes the HBV reverse transcriptase expressed in E. coli.

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Activation of the Vibrio vulnificus cadBA Operon by Leucine-Responsive Regulatory Protein is Mediated by CadC

  • Rhee, Jee-Eun;Kim, Kun-Soo;Choi, Sang Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1755-1761
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    • 2008
  • The present study revealed that Lrp, a leucine-responsive regulatory protein, is involved in the regulation of cadBA transcription through activation of $P_{cadBA}$. The influence of Lrp on $P_{cadBA}$ was mediated by CadC, and thereby, CadC was able to compensate for the lack of Lrp in the activation of $P_{cadBA}$. Western blot analyses and EMSA demonstrated that the cellular level of CadC was not significantly affected by Lrp, and that Lrp exerted its effect by directly binding to $P_{cadBA}$. These combined results suggested that CadC and Lrp function cooperatively to activate the $P_{cadBA}$ rather than sequentially in a regulatory cascade.

Comparison of tdcA Expression Between Escherichia coli and Salmonella enterica Serovar Typhimurium

  • Kim, Min-Jeong;Lim, Sang-Yong;Ryu, Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.252-255
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    • 2011
  • Both Escherichia coli (E. coli) and Salmonella enterica serovar Typhimurium (S. Typhimurium) have a tdc operon that encodes enzymes involved in a metabolic pathway for the degradation of L-serine and L-threonine. However, S. Typhimurium does not have the tdcR gene, which is a positive regulator in E. coli. In the present study, transcriptional analysis revealed that tdcA expression in E. coli is higher under anaerobic than aerobic growth conditions, but the opposite is true in S. Typhimurium. Interestingly, a tdcR mutant strain of E. coli showed a similar expression pattern to that observed in S. Typhimurium and was also induced by anaerobic shock. These results suggest that the induction of tdcA expression by anaerobic conditions is observable when tdcA expression is low owing to the absence of TdcR.

Characterization of Protein Factor Regulating the Superoxide-Sensor SoxR in Escherichia coli

  • Koo, Mi-Sun;Rah, So-Yeon;Lee, Kang-Lok;Roe, Jung-Hye
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.64-64
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    • 2001
  • Escherichia coli has developed soxRS regulon to defend against toxicity of superoxide radical. SoxR, superoxide sensor, is oxidized by superoxide-generating agents or nitric oxide and oxidized SoxR activates the transcription of soxS gene. In order to find out the trans-acting factors regulating SoxR activity in vivo, soxS::lacZ single copy operon fusion construct was prepared and random Tn10 insertional mutatons were performed.(omitted)

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