• 제목/요약/키워드: lac I

검색결과 112건 처리시간 0.024초

Toxic Mechanism of Atrazine in Transgenic Mutagenesis System using Big $Blue^{\textregistered}}$Rat2 lacI Transgenic Fibrohlasts And Hormonal Disturbances in vitro

  • Kim, Youn-Jung;Lee, Kilchul;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2001년도 춘계심포지움 및 학술발표회
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    • pp.128-128
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    • 2001
  • Atrazine, one of herbicide widely used in agriculture, is classified as a possible human carcinogen (2B group) that may cause breast and ovarian cancers by IARC, and is known as an endocrine disrupter. Atrazine has been subjected to broad ranges of genotoxicity tests wi th predominantly negative results, but reported conflicting results across two or more independent tests as well. (omitted)

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TOXIC MECHANISM OF ATRAZINE IN TRANSGENIC MUTAGENESIS SYSTEM USING BIG BLUE$\circledR$ RAT2 lacI TRANSGENIC FIBROBLASTS AND HORMONAL DISTURBANCES IN VITRO

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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    • pp.183-183
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    • 2001
  • Atrazine, one of herbicide widely used in agriculture, is classified as a possible human carcinogen (2B group) that may cause breast and ovarian cancers by IARC, and is known as an endocrine disruptor. Atrazine has been subjected to broad ranges of genotoxicity tests with predominantly negative results, but reported conflicting results across two or more independent tests as well. This fact indicates that a more comprehensive genotoxicity assessment needs for atrazine.(omitted)

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플라스미드 pKM101 과 pSL4 의 muc 유전자의 발현에 관한 연구 (Expression of mue Gene on Plasmid pKM101 and pSL4)

  • 전홍기;황유경;이상률;백형석
    • 미생물학회지
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    • 제30권5호
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    • pp.371-376
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    • 1992
  • 플라스미드 pKM101 과 이의 돌연변이체 pSL4 는 UV 및 MMS 에 대해 높은 치사 저항성과 돌연변이률을 나타낸다. pKM101 과 pSLA 의 mucB 유전자의 일부분과 mucA 유전자를 lacZ fusion 벡터인 pMC874 에 subcloning 시켜 플라스미드 pBH31 과 pBH30 을 선별하였고 이 플라스미드들을 $recA^{+}lexA^{-}$, $recA^{-}와lexA^{+}$, 균주에 각각 도입하였다. $recA^{+}lexA^{+}$ 균주에서 pSLA 의 muc 유전자를 포함하는 pBH30 의 $\beta$-galactosidase활성은 pKM101 의 muc 유전자가 연결된 pBH31 보다 높았고 $recA^{-}$$lexA^{-}$ 돌연변이주에서는 UV 조사의 유무에 관계없이 $\beta$-galactosidase를 유도하지 못하였지만 $recA^{-}$ 균주에서는 UV 조사를 하지 않았을 때 pBH30 의 $\beta$-galactosidase가 pBH31 보다 약간 높게 유도되었다. 이러한 결과는 pKM101 과 pSLA 의 기능적 차이가 두 플라스미드의 Muc 단백질의 구조적 차이라고 생삭할 수 있지만, muc 유전자의 조절부위가 돌연변이되너 LexA repessor 가 작용하는 부위의 변화에 의한 것이라고도 생각햐ㄹ 수 있었다. 또한, umuC-lacZ 유전자를 가지는 pBH100 를 construction 하여 umu oeron 의 발현은 UV 조사에 의해 유도되며 recA 와 lexA 유전자에 의해 조절됨을 알 수 있었다.

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Genes of Rhodobacter sphaeroides 2.4.1 Regulated by Innate Quorum-Sensing Signal, 7,8-cis-N-(Tetradecenoyl) Homoserine Lactone

  • Hwang, Won;Lee, Ko-Eun;Lee, Jeong-Kug;Park, Byoung-Chul;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • 제18권2호
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    • pp.219-227
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    • 2008
  • The free-living photoheterotrophic Gram-negative bacterium Rhodobacter sphaeroides possesses a quorum-sensing (QS) regulatory system mediated by CerR-CerI, a member of the LuxR-LuxI family. To identify the genes affected by the regulatory system, random lacZ fusions were generated in the genome of R. sphaeroides strain 2.4.1 using a promoter-trapping vector, pSG2. About 20,000 clones were screened and 23 showed a significantly different level of ${\beta}$-gal activities upon the addition of synthetic 7,8-cis-N-tetradecenoyl-homoserine lactone (RAI). Among these 23 clones, the clone showing the highest level of induction was selected for further study, where about a ten-fold increase of ${\beta}$-gal activity was exhibited in the presence of RAI and induction was shown to be required for cerR. In this clone, the lacZ reporter was inserted in a putative gene that exhibited a low homology with catD. A genetic analysis showed that the expression of the catD homolog was initiated from a promoter of another gene present upstream of the catD. This upstream gene showed a strong homology with luxR and hence was named qsrR (quorum-sensing regulation regulator). A comparison of the total protein expression profiles for the wild-type cells and qsrR-null mutant cells using two-dimensional gel electrophoresis and a MALDI-TOF analysis allowed the identification of sets of genes modulated by the luxR homolog.

Enterobacter aerogenes 의 phoA 유전자 Promoter를 이용한 인 제한환경에서 발현하는 벡터 구축 (Construction of the Phosphate-Limitation Inducible Expression Vector Containing the phoA Promoter of Enterobacter aerogenes)

  • 장화형;고병훈;박신영;이성호;김성진;임유정;한갑진;김영호;이영근
    • 미생물학회지
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    • 제38권4호
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    • pp.318-321
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    • 2002
  • 토양 등의 인 제한환경에서 특이적으로 발현하는 벡터를 구축하기 위해서 Enterobacter areogenes의 phoA 유전자의 promoter가 든 pEAAP를 구축하였다. pEAAP는 pET-22b(+)을 BglII와 XbaI으로 절단하여 T7 promoter와 lac operator를 제거하고pho box가 포함된 phoA promoter를 삽입하여 구축하였다. pEAAP가 인 제한 환경에서 특이적으로 발현되는지 조사하고자 Bacillus subtillis var. amyloliquefaciens (KCTC 8913P)의 Phytase유전자인 Bsa-phy1을 도입한 pEAPHY1을 구축하였다. CK-PHY1 (pEAPHY1을 도입한Escherichia coli JM109)는 인 제한 환경에서 41 kD)의 Bsa-Phy1을 발현하였다. 또한, CK-PHY1은 phytate를 유일한 인산원으로 첨가된 고체배지에서 phytate를 분해하여 투명대를 형성하였다.

A closer look at the structure and gamma-ray shielding properties of newly designed boro -tellurite glasses reinforced by bismuth (III) oxide

  • Hammam Abdurabu Thabit;Abd Khamim Ismail;N.N. Yusof;M.I. Sayyed;K.G. Mahmoud;I. Abdullahi;S. Hashim
    • Nuclear Engineering and Technology
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    • 제55권5호
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    • pp.1734-1741
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    • 2023
  • This work presents the synthesis and preparation of a new glass system described by the equation of (70-x) B2O3-5TeO2 -20SrCO3-5ZnO -xBi2O3, x = 0, 1, 5, 10, and 15 mol. %, using the melt quenching technique at a melting temperature of 1100 ℃. The photon-shielding characteristics mainly the linear attenuation coefficient (LAC) of the prepared glass samples were evaluated using Monte Carlo (MC) simulation N-particle transport code (MCNP-5) at gamma-ray energy extended from 59 keV to 1408 keV emitted by the radioisotopes Am-241, Ba-133, Cs-137, Co-60, Na-22, and Eu-152. Furthermore, we observed that the Bi2O3 content of the glasses had a significantly stronger impact on the LAC at 59 and 356 keV. The study of the lead equivalent thickness shows that the performance of fabricated glass sample with 15 mol.% of Bi2O3 is four times less than the performance of pure lead at low gamma photon energy while it is enhanced and became two times lower the perforce of pure lead at high energy. Therefore, the fabricated glasses special sample with 15 mol.% of Bi2O3 has good shielding properties in low, intermediate, and high energy intervals.

Transcriptional Regulation of the Schizosaccharomyces pombe Gene Encoding Glutathione S-Transferase I by a Transcription Factor Pap1

  • Kim Hong-Gyum;Kim Byung-Chul;Kim Kyunghoon;Park Eun-Hee;Lim Chang-Jin
    • Journal of Microbiology
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    • 제42권4호
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    • pp.353-356
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    • 2004
  • In a previous study, a gst gene was isolated from the fission yeast Schizosaccharomyces pombe. This gene was dubbed gstI, and was characterized using the gstI -lacZ fusion plasmid pYSH2000. In this work, four additional fusion plasmids, pYSHSDl, pYSHSD2, pYSHSD3 and pYSHSD4, were constructed, in order to carry (respectively) 770, 551, 358 and 151 bp upstream regions from the translational initiation point. The sequence responsible for induction by aluminum, mercury and hydrogen peroxide was located in the range between -1,088 and -770 bp upstream of the S. pombe gstI gene. The same region was identified to contain the nucleotide sequence responsible for regulation by Papl, and has one puta­tive Papl binding site, TTACGTAT, located in the range between $-954\~-947$ bp upstream of the gstI gene. Negatively acting sequences are located between -1,088 and -151 bp. These findings imply that the Papl protein is involved in basal and inducible transcription of the gstI gene in the fission yeast S. pombe.

Bacillus stearothermophilus 의 내열성 시티딘/디옥시시티딘 디아미나제를 코드하는 cdd 유전자의 클로닝 (Molecular Cloning of Bacillus stearothermophilus cdd Gene Encoding Thermostable Cytidine/Deoxycytidine Deaminase)

  • Soo, Chang-Jong;Song, Bang-Ho;Kim, Jong-Guk;Hong, Soon-Duck
    • 한국미생물·생명공학회지
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    • 제17권4호
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    • pp.334-342
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    • 1989
  • Bacillus stearothermophilus의 cytidine deaminase (cytidine/2'-deoxycytidine aminohydrolase:EC 3.5.4.5)를 코딩하는 cdd 유전자를 E. coli cdd$^-$ 결손변이주를 cloning host로 하여 3-10Kbp의 B. stearothermophilus DNA 단편으로부터 shot gun 법으로 클로닝하였다. 고 복제수 플라스미드 pBR322 의 PstI 부위에 3.0Kb의 B. stearothermophilus DNA 단편을 함유한 pJSC101이 cdd$^+$와 tetracy-line 내성으로서 cloning되었으며, 이어서, 결실 및 subcloning을 연속 수행한 결과 약 1.35kbp의 Eco RI$_1$/PstI$_2$단편이 동일 부위의 pBR322에 삽입된 cdd 양성의 pJSC201을 얻었다. Mini 세포 실험결과, 이 단편에서 합성되는 polypeptide는 약 33 KDa이었기에 이 polypeptide가 cytidine deaminase 로 추정되었다. 또한 이 단편에 함유한 550bp의 EcoRI/AvaI 부분을 lacZ 프로모터 영역에 삽입한 경우 프로모터 활성을 나타내었기에 이 단편의 Eco RI 부위에서 PstI부위로 cdd 유전자가 전사됨을 알 수 있었다. B. subilis와 E. coli에서 발현이 가능한 shuttle vector에 cdd가 함유된 단편을 삽입한 후 이를 양세포에서 동시 발현시켰을 때 B. subtilis에서 발현시킨 경우가 E. coli에서 보다높은 cytidine deaminase 활성을 나타내었으며 이 유전자는 B. subtilis 에서도 E. coli에서와 같이 안정하게 유지됨을 알 수 있었다.

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유전공학 기법을 이용한 새로운 당뇨병 치료제 개발 연구

  • 남두현
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.266-266
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    • 1994
  • $B^{30}$ 위치에 homoserine이 치환된 사람 insulin 유사체 ($B^{30}$ -homoserine) insulin을 생산하기 위해, insulin의 B 사슬 유전자에 A 사슬 유전자를 직접 연결한 insulin 유전자를 설계하였다. 이 유전자는 10개의 oligonucleotide로 나누어 합성하여 T4 DNA ligase로 결합시킨 후, pUC19 plasmi의 polylinker 영역에 삽입하였다. 이 유전자의 발현을 높이기 위해 이 유전자는 다시 tac promoter의 지배를 받는 lacZ 유전자의 Cia I 또는 Hpa I 제한부위에 도입하여 융합시켰다. 이렇게 구축된 운반체 pTBA나 pKBA를 Escherichia coli JM103 균주에 형질도입시킨 후, 이를 4시간 배양한 후 0.05mM이상의 isopropyl-$\beta$-D-thiogalactopyranoside (IPTG)를 배지에 공급해 주고 2시간 더 배양하였을 때 유전자 발현이 잘 유도되어짐을 알 수 있었다. 이 때 생산된 insulin 전구체들은 세포내 불온성인 inclusion body로 축적되어지는 것을 관찰하였으며, 그 생산량은 세포내 전체 단백질량의 30%에 달하였다.

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Post-pillars design for safe exploitation at Trepça hard rock mine (Kosovo) based on numerical modeling

  • Ibishi, Gzim;Genis, Melih;Yavuz, Mahmut
    • Geomechanics and Engineering
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    • 제28권5호
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    • pp.463-475
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    • 2022
  • In the mine exploitation stage; one of the critical issues is the stability assessment of post-pillars. The instability of post-pillars leads to serious safety hazards in mining operations. The focus of this study is to assess the stability of post-pillars in the 130# stope in the central ore body at Trepça hard rock mine by employing both conventional (i.e., critical span curve) and numerical methods (i.e., FLAC3D). Moreover, a new numerical based index (i.e., Pillar Yield Ratio-PYR) was proposed. The aim of PYR index is to determine a border line between stable, potentially unstable, and failure state of post-pillars at a specific mine site. The critical value of pillar width to height ratio is 2.5 for deep production stopes (e.g., > 800 m). Results showed that pillar size, mining height and mining depth significantly have affected the post-pillar stability. The reliability of numerical based index (i.e., PYR) is verified based on empirical underground pillar stability graph developed by Lunder, 1994. The proposed pillar yield ratio index and pillar stability graph can be used as a design tool in new mining areas at Trepça hard rock mine and for other situations with similar geotechnical conditions.