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대장균을 이용한 Phenylalanine 생산에 있어서 온도조절형 발현 Vector의 안정성 (Phenotypic Stability of a Temperature-Controllable Expression Vector on Phenylalanine Production by Escherichia coli)

  • 강상모;박인숙
    • 한국미생물·생명공학회지
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    • 제19권5호
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    • pp.433-438
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    • 1991
  • Phenylalanine 고생산용 plasmid pSY130-14는 $\lambda$-phage 유래 온도감수성을 가지고 있으므로 $cI_{857}$ repressor와 PL과 PR을 온도를 올려 phenylalanine의 생산을 유도한다. pSY130-14를 갖는 E.coli AT2471은 kanamycin 무첨가, $38.5^{\circ}C$, 48시간에서 약 30%로 plasmid가 없어지며 첨가에서 안정성이 떨어졌다가 배양시간과 더불어 올라갔는데, 이것은 생육에 피요한 kanamycin gene과 ori만이 남는 것으로 생각된다.

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Effects of Vitamin C and Vitamin E Supplementation on Anti-oxidative System of the Smokers and Non-smokers

  • Kim Hyun Ah;Min Hye Sun;Ha Ae Wha;Hyun Hwa Jin;Lee Hong Mie;Ro Man Soo;Song Kyung Hee
    • Journal of Community Nutrition
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    • 제6권3호
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    • pp.146-154
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    • 2004
  • This study compared intake of vitamins and antioxidant nutritional status of smokers and nonsmokers, and the effect of supplementation of vitamin C and vitamin E on antioxidant system of smokers and nonsmokers. Subjects were 30 smokers and 30 non-smokers of male university students. They were divided into groups of 10 subjects each one with supplementation for 4 weeks, to investigate the effects of supplementation. Smokers were divided into vitamin C supplement group, vitamin E supplement group and vitamin C and vitamin E combination supplement group, and so were nonsmokers. The supplementation of vitamin C was 500mg and vitamin E was 400IU. There was no significant difference of antioxidant vitamin intakes between smokers and non-smokers, and plasma concentration of vitamin C in smokers was lower than non-smokers (p<0.05). Blood pressure was higher in smokers (p<0.05), with no difference in blood glucose levels, methemoglobin and TBARS, but SOD activity was significantly higher in non-smokers (p<0.001). Vitamin C supplementation resulted in a significant decrease of diastolic blood pressure (p<0.01), systolic blood pressure (p<0.001) and methemoglobin (p<0.001) in smokers. Also a significant decrease of diastolic blood pressure (p<0.05), systolic blood pressure (p<0.05), blood glucose (p<0.05), methemoglobin (p<0.001) and TBARS (p<0.05), with significant increase of SOD activity (p<0.001) were found in vitamin E supplement group. In vitamin C and vitamin E combination supplement group, there was a significant decrease of diastolic blood pressure (p<0.05), methemoglobin (p<0.01) and TBARS (p<0.05). In non-smokers, methemoglobins (p<0.001) of vitamin C supplement group and vitamin E supplement group decreased, and diastolic pressure (p<0.05), methemoglobin (p<0.01) and TBARS (p<0.05) significantly decreased in vitamin C and vitamin E combination supplement group. These results indicated better efficacy of antioxidant supplementation in smokers than in nonsmokers, suggesting that the supplementation of vitamin C and vitamin E might decrease the oxidative stress and various risk factors of smoking-related diseases.

P.T.C.R.-N.T.C.R. 자기를 이용한 저항체에 관한 연구 (A Study on the P.T.C.R-N.T.C.R. Ceramic Resister)

  • 안영필;이병하;김일기
    • 한국세라믹학회지
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    • 제18권2호
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    • pp.119-125
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    • 1981
  • The thermostable resistor was made by using P.T.C.R-N.T.C.R. ceramics. In this study $BaTiO_3$ type P.T.C.R. ceramics and $SnO_2-Sb_2O_3$ type N.T.C.R. ceramics were cocrystallized in one body and electrical resistance was stable under the temperature variation between $25^{\circ}C$-40$0^{\circ}C$. The ceramics showed the most stable electrical resistance for the mal variation when the composition was 0.6P.T.C.R. $BaTiO_3$,$ 0.4SnO_2$ and 0.01 $Sb_2O_3$.

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Synthesis and Reactivity of the Pentacoordinate Organosilicon and -germanium Compounds Containing the C,P-Chelating ο-Carboranylphosphino Ligand [ο-C2B10H10PPh2-C,P](CabC,P

  • Lee, Tae-Gweon;Kim, Sang-Hoon;Kong, Myong-Seon;Kang, Sang-Ook;Ko, Jae-Jung
    • Bulletin of the Korean Chemical Society
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    • 제23권6호
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    • pp.845-851
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    • 2002
  • The synthesis of the intramolecular donor - stabilized silyl and germyl complexes of the type ($Cab^c.p) MMe_2X$ (2a:M=Si, X=Cl;2b;M= Ge, X=Cl;2e;M=Si,X=H) was achieved by the reaction of $LiCab^c,p$ (1) with $Me_2SiClX$ and $Me_2GeCl_2$ respectively. The intramolecular M←P interacion in 2a-2c is provided by $^1H$, $13^C.$, $31^P$ and $29^Si$ NMR spectroscopy. The salt elimination reactions of dichlorotetramethyldisilane and -digermane with 1 afforded the $bis(\sigma-carboranylphosphino)disilane$ and disgermane [$(Cab^C.P)MMe_2]_2(4a;M$ = Si;4b: M=Ge). The oxidative addition reaction of 4a-4b with $pd_2(dba)_3CHCl_3afforded$ the bis(silyl)-and bis(germyl)-palladium complexes. The chloro-bridged dipalladium complexes were obtained by the reaction of 2a-2b with $pd_2(dba)_3CHCl_3$ The crystal structures of 5a and 7b were determined by X-ray structural studies.

Effects of Enhanced pCO2 and Temperature on Reproduction and Survival of the Copepod Calanus sinicus

  • Kang, Hyung-Ku;Lee, Chang-Rae;Kim, Dongseon;Yoo, Sinjae
    • Ocean and Polar Research
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    • 제38권4호
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    • pp.303-314
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    • 2016
  • We tested the combined effects of increased partial pressure of $CO_2$ ($pCO_2$) and temperature on the reproduction and survival of the copepod Calanus sinicus from Asan Bay, the Yellow Sea under laboratory conditions to understand the impact of acidification on copepods. Egg production rate, survival rate, and fecal pellet production of C. sinicus were not affected by 1305 ppm $pCO_2$ or with combined treatments of temperature and $pCO_2$, including $8^{\circ}C$ and 289 ppm $pCO_2$ (ambient), $8^{\circ}C$ and 753 ppm $pCO_2$ (high $pCO_2$), $12^{\circ}C$ and 289 ppm $pCO_2$ (high temperature), and $12^{\circ}C$ and 753 ppm $pCO_2$ (greenhouse), for 5 or 10 d of exposure. However, egg hatching success of C. sinicus decreased significantly in the greenhouse treatment compared with the ambient or the high $pCO_2$ treatments. These results suggest that a combined treatment ($pCO_2$ and temperature) affected egg viability more than a single treatment($pCO_2$).

The Bacteriophage λ DNA Replication Protein P Inhibits the oriC DNA- and ATP-binding Functions of the DNA Replication Initiator Protein DnaA of Escherichia coli

  • Datta, Indrani;Sau, Subrata;Sil, Alok Kumar;Mandal, Mitai C.
    • BMB Reports
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    • 제38권1호
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    • pp.97-103
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    • 2005
  • Under the condition of expression of $\lambda$ P protein at lethal level, the oriC DNA-binding activity is significantly affected in wild-type E. coli but not in the rpl mutant. In purified system, the $\lambda$ P protein inhibits the binding of both oriC DNA and ATP to the wild-type DnaA protein but not to the rpl DnaA protein. We conclude that the $\lambda$ P protein inhibits the binding of oriC DNA and ATP to the wild-type DnaA protein, which causes the inhibition of host DNA synthesis initiation that ultimately leads to bacterial death. A possible beneficial effect of this interaction of $\lambda$ P protein with E. coli DNA initiator protein DnaA for phage DNA replication has been proposed.

Corynebacterium glutamicum-Escherichia coli Shuttle Vector 개발과 C.glutamicum 의 Homoserine Dehydrogenase Gene Cloning (Construction of a Corynebacteriurn glutarnicum-Escherichicr coli Shuttle Vector and Cloning the Homoserine ehydrogenase Gene from C. glutamicum)

  • 최신건;박종현;신현경
    • 한국미생물·생명공학회지
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    • 제19권1호
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    • pp.31-36
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    • 1991
  • Tn5의 kanamycin 저항성 유전자를 가진 pBEL1 plasmid와 C.glutamicum cryptic plasmid인 pSR1으로 7.5kb의 새로운 plasmid를 만든 후, 이를 pCE1301이라 명명하였다. 이 pCE1301은 PEG1301은 PEG-protoplast법으로 C.glutamicum을 형질전환하였을 때 효율이 약 $3.0\times 10^3$형질전환제/$\mu g$이었으며 SalI과 EcoRI 제한효소 절단부위가 1개씩이었다. 또 Km이 없는 배지에서 25세대까지 안정하게 유지되었으며 B.flavum, E.coli에서 복제되었다. 이 pCE1301을 이용하여 C.glutamicum 의 homoserine dehydrogenase 유전자를 cloning하였다.

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Functional Characterization of Pharmcogenetic Variants of Human Cytochrome P450 2C9 in Korean Populations

  • Cho, Myung-A;Yoon, Jihoon G.;Kim, Vitchan;Kim, Harim;Lee, Rowoon;Lee, Min Goo;Kim, Donghak
    • Biomolecules & Therapeutics
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    • 제27권6호
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    • pp.577-583
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    • 2019
  • Human cytochrome P450 2C9 is a highly polymorphic enzyme that is required for drug and xenobiotic metabolism. Here, we studied eleven P450 2C9 genetic variants-including three novel variants F69S, L310V, and Q324X-that were clinically identified in Korean patients. P450 2C9 variant enzymes were expressed in Escherichia coli and their bicistronic membrane fractions were prepared The CO-binding spectra were obtained for nine enzyme variants, indicating P450 holoenzymes, but not for the M02 (L90P) variant. The M11 (Q324X) variant could not be expressed due to an early nonsense mutation. LC-MS/MS analysis was performed to measure the catalytic activities of the P450 2C9 variants, using diclofenac as a substrate. Steady-state kinetic analysis revealed that the catalytic efficiency of all nine P450 2C9 variants was lower than that of the wild type P450 2C9 enzyme. The M05 (R150L) and M06 (P279T) variants showed high $k_{cat}$ values; however, their $K_m$ values were also high. As the M01 (F69S), M03 (R124Q), M04 (R125H), M08 (I359L), M09 (I359T), and M10 (A477T) variants exhibited higher $K_m$ and lower $k_{cat}$ values than that of the wild type enzyme, their catalytic efficiency decreased by approximately 50-fold compared to the wild type enzyme. Furthermore, the novel variant M07 (L310V) showed lower $k_{cat}$ and $K_m$ values than the wild type enzyme, which resulted in its decreased (80%) catalytic efficiency. The X-ray crystal structure of P450 2C9 revealed the presence of mutations in the residues surrounding the substrate-binding cavity. Functional characterization of these genetic variants can help understand the pharmacogenetic outcomes.

김치유산균용 발현벡터 pSJE6c 개발과 이를 이용한 외래 유전자 발현 (Development of pSJE6c, an Expression Vector for Kimchi Lactic Acid Bacteria, and Heterologous Gene Expression Using the Vector)

  • 이강욱;박지영;이지연;이황아;백창운;조현덕;김주연;권건희;천지연;김정환
    • 한국미생물·생명공학회지
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    • 제37권4호
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    • pp.389-398
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    • 2009
  • 본 연구실에서 개발한, 김치에서 분리한 Leu. mesenteroides SY2 유래 pFMBL1 을 바탕으로 구축한 셔틀벡터인, pSJE[7]를 외래유전자 발현에 적합하게 개량한 발현벡터를 구축하였다. Lactococcus lactis LM0230에서 분리한 프로모터 P6C를 pSJE에 도입하였다. P6C 염기서열을 지닌 oligonucleotide 쌍을 따로 제조한 후 annealing을 통해 짧은 DNA 단편을 얻어서 제한효소 처리후 pSJE에 도입하여 pSJE6c를 구축하였다. PSJE6c 효능 검증을 위해서 외래 유전자인 aga와 lacZ를 각각 pSJE6c에 도입하였다. P6C 프로모터와 비교를 위해 고유 프로모터를 지닌 유전자들도 각각 pSJE에 도입하였다. 재조합 plasmid들을 electroporation 방법으로 Lactobacillus brevis 2.14 균주에 도입하고 재조합균주들의 생육곡선과 효소역가 그리고 slot blot으로 전사체 농도를 측정하였다. 결과를 보면 PSJE6c에 클로닝 된 유전자들이 pSJE상의 유전자보다 효소역가들이 약 1.5배에서 2배 정도로 높았다. 전사체 농도 측정 결과도 pSJE6c 들에서 더 많은 전사체가 생성됨을 보여주었다. 이상 결과들은 효율적인 발현벡터들의 사용을 통해서 김치유산균에서 외래유전자 발현 효율을 높일 수 있음을 보여준다.

집단체조프로그램이 여성노인의 신체조성, 혈청지질에 미치는 효과 (The Effects of Group Exercise on Fat Distribution, Lipid Profiles in Elderly Women)

  • 이경혜;강현숙
    • 재활간호학회지
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    • 제8권1호
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    • pp.13-19
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    • 2005
  • Purpose: This study was aimed to identify the effects of group exercise on physiological functions in Elderly. Method: The design of research was one group pretest-posttest design. The subjects were sixty two elderly women persons over sixty years old to live in C-GUN, The exercise period was 12-weeks and frequency was three times per week. the exercise duration was sixty minites. In order to evaluate the effect of exercise, we measured %body fat, fat mass(FM), fat free mass(FFM), body mass index(BMI), body metabolic rate(BMR), Total Cholesterol(TC), Triglyceride(TG), high density lipoprotein-c(HDL-C), low density lipoprotein-c(LDL-C) before and after the 12 week's group exercise program. The data analyzed by the paired t-test using SPSS package. Result: There was significant difference in FFM(p=.000), BMR(p=.000), HDL-C(p=.010). There was no significant difference in weight(p=.122), %body fat(p=.062),, FM(p=.704), BMI(p=.136), TC(p=.125), TG(p=.064), LDL-C(p=.575) Conclusion: This group exercise for elderly women can be recommended as an effective nursing intervention for elderly women in a community.

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