• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.71-80
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• 2005

Effects of low temperature ($8^{\circ}C$) on the hydraulic conductivity of young roots of a chilling-sensitive (cucumber; Cucumis sativus L.) and a chilling-resistant (figleaf gourd; Cucurbita ficifolia Bouche) crop have been measured at the levels of whole root systems (root hydraulic conductivity, $Lp_r$) and of individual cortical cells (cell hydraulic conductivity, Lp). In figleaf gourd, there was a reduction only in hydrostatic $Lp_r$ but not in osmotic $Lp_r$ suggesting that the activity of water channels was not much affected by low root temperature (LRT)treatment in this species. Changes in cell Lp in response to chilling and recovery were similar asroot level, although they were more intense at the root level. Roots of figleaf gourd recovered better from LRT treatment than those of cucumber. In figleaf gourd, recovery (both at the root and cell level) often resulted in Lp and $Lp_r$ values which were even bigger than the original, i.e. there was an overshoot in hydraulic conductivity. These effects were larger forosmotic (representing the cell-to-cell passage of water) than for hydrostatic $Lp_r$. After a short term (1 d) exposure to $8\;^{\circ}C$ followed by 1 d at $20\;^{\circ}C$, hydrostatic $Lp_r$ of cucumber nearly recovered and that of figleaf gourd still remained higher due to the overshoot. On the contrary, osmotic $Lp_r$ and cell Lp in both species remained high by a factor of 3 as compared to the control, possibly due to an increased activity of water channels. After pre-conditioning of roots at LRT, increased hydraulic conductivitywas completely inhibited by $HgCl_2$ at both the root and cell levels. Different from figleaf gourd, recovery from chilling was not complete in cucumber after longer exposure to LRT. It is concluded that at LRT, both changes in the activity of aquaporins and alterations of root anatomy determine the water uptake in both species. To better understand the aquaporin function in plants under various stress conditions, we examined the transgenic Arabidopsisand tobacco plants that constitutively overexpress ArabidopsisPIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates were found between the transgenic and wild-type plants under favorable growth conditions. By contrast, overexpression of PIP1;4 or PIP2;5 had a negative effect on seed germination and seedling growth under drought stress, whereas it had a positive effect under cold stress and no effect under salt stress. Measurement of water transport by cell pressure probe revealed that these observed phenotypes under different stress conditions were closely correlated with the ability of water transport by each aquaporin in the transgenic plants. Together, our results demonstrate that PIP-type aquaporins play roles in seed germination, seedling growth, and stress response of Arabidopsis and tobacco plants under various stress conditions, and emphasize the importance of a single aquaporin-mediated water transport in these cellular processes.

• 한국미생물·생명공학회지
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• 제13권4호
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• pp.349-354
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• 1985

Plasmid pBR322와 runaway Plasmid pSY343을 vector로 사용하여 E. stearothermophilus IAM 11062내의 $\alpha$-amylase 유전자를 E. coli내에 클로닝 하였다. 이때 얻어진 $\alpha$-amylase유전자는 제한효소 Hind III의 말단을 갖고 있는 4.7kb의 크기였으며 E. coli내에서 이들 유전자는 비교적 안정적 있게 유지되고 발현되었다. 재조합 $\alpha$-amylase유전자가 클로닝된 E. coli는 B. stearothermophilus IAM 11062보다 3배의 $\alpha$-amylase를 더 많이 생성하였다. EDTA를 사용한 osmotic shock 방법에 의하여 E. coli내에서 생성된 $\alpha$-amylase는 그 효소 생성량의 75%정도가 periplasm에 존재함이 밝혀졌다. 재조합된 $\alpha$-amylase 유전자에 의해서 E. coli에서 생성된 $\alpha$-amylase는 최적 작용온도가 55$^{\circ}C$로서 이들의 열안정성과 분자량(61,000)도 B. stearothermophilus IAM 11062의 $\alpha$-amylase와 거의 동일하게 나타나 E. coli와 B. stearothermophilus IAM 11062에서 생성된 $\alpha$-amylase는 효소학적 성질이 같음을 보여주었다.

• 미생물학회지
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• 제34권3호
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• pp.154-161
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• 1998

자연미생물, KP964에 luxAB 유전자를 주입하여 발광기능을 부여함으로써 그들의 발광정도가 스트레스에 대한 적응, 생존 및 활성도를 나타내는 색인이 될 수 있는가, 그리고 특히 non-culturable but viable(NCBV) 상태로 존재하는 미생물의 활성에 대한 정보를 제공할 수 있는가를 알아보았다. 영양결핍조건 하에서 그들의 총 세균수, 배양가능세균(colony-forming-unit; CFU)수, viable cell 수, 그리고 발광정도(relative light unit; RLU)의 변화를 조사하였다. 형광현미경으로 확인된 총 수는 변화가 없었으며, 첨가된 yeast extract에 의해 성장하는 viable cell 수는 완만하게 감소하였으나 CFU 수는 급격히 줄어들어 37일 째 이후에는 탐지한계 이하로 감소하였다. RLU의 경우, 처음 7일의 영양결핍동안 0.016%까지 감소하였으며, 그 이후에는 탐지한계수준 이하로 검출되었다. 아울러 스트레스를 받는 4시간 동안 CFU수와 RLU의 관계를 알아보기 위하여 발광 KP964 균주에게 독성유해물질, acid shock, osmotic shocks를 접촉시킨 후 시간별로 CFU와 RLU를 측정한 결과, KP964 CFU 당 RLU는 9.1-26% 까지 감소하였다. 이는 스트레스의 종류에 따라 접촉시간이 길수록 CFU 당 나오는 빛의 양이 줄어듬을 보인 것이다. 이러한 결과는 영양결핍조건 시 RLU의 감소율이 CFU의 감소율보다 더 크며 영양결핍 이외의 다른 스트레스 조건 하에서도 RLU의 감소율과 CFU의 감소율의 연관성을 찾을 수 없음을 보이므로, 발하는 빛의 정량만으로는 NCBV 상태는 물론 배양가능한 세균의 양이나 활성을 나타내지 못하는 듯 하였다. 그러나 스트레스를 받은 KP964의 활성 정도와 잠재발광능력과의 관계는 좋은 연관성을 보여주었다. 즉, p-xylene, hypo-osmotic stress, 또는 영양결핍에 노출된 세균으로부터 각 스트레스를 제거하였을 시 나타나는 발광증가 유형을 조사하였는데, 증가가 시작하기까지의 lag period는 각각 32, 26, 22분으로 나타났다. 즉 미생물에 강력한 스트레스로 작용한 경우일수록 lag time이 길고, 또한 발광의 증가율이 느리며 도달하는 최대 RLU값도 낮음이 관찰되었다. 따라서 스트레스 하의 세균으로부터 직접적인 RLU의 측정보다는 이들이 스트레스 조건으로부터 재성장 또는 소생되는 과정에서의 잠재 발광능력의 분석이 자연세균의 적응, 활성 및 생존의 정도를 나타내주었다.

• 한국버섯학회지
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• 제20권3호
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• pp.178-182
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• 2022

버섯의 오랜 역사에도 불구하고 버섯의 유전적 기능과 분자유전학을 응용한 신품종 개발에 대한 연구는 크게 부족한 상황이다. 그러나 최근 유전자 가위인 CRISPR/Cas를 이용한 새로운 유전자 교정 기술이 개발됨에 따라 버섯 연구에서 이 기술을 이용한 다양한 시도가 이루어지고 있다. 특히 선택의 용이성을 위해 외래 유전자 삽입 없이도 고효율로 유전자 편집이 가능한 RNPs를 활용한 연구가 활발히 진행되고 있다. 그러나 RNPs는 원형질체의 세포막을 통과하기에 Cas9이 너무 거대하고 guide RNA가 쉽게 파괴된다는 단점을 가지고 있다. 이러한 단점을 극복하기 위하여 세포막 통과에 용이한 미네랄 성분인 CaP와 PAA를 조합하여 Nanoparticle을 형성함으로써 극복하고자 했다. 표고버섯 단핵 균주인 산조705-13을 이용하여 원형질체 분리에 적합한 Osmotic buffer를 찾기 위하여 0.6M과 1.2M의 Sucrose, Sorbitol, Mannitol, KCl을 처리하였고 그 결과 0.6M Sucrose가 가장 적합한 osmotic buffer임을 확인하였다. 또한 CaP으로 RNPs와 Nanoparticle 복합체를 형성하고 이 복합체가 RNase A로부터 RNPs의 기능을 온전히 보호하는 것을 확인할 수 있었다.

• The Korean Journal of Physiology
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• 제17권2호
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• pp.135-142
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• 1983

$PGE_2$ 및 $PGF_{2{\alpha}}$가 용혈 및 적혈구막 절편에서 $Ca^{++}$ 결합에 미치는 영향을 관찰하여 다음과 같은 결론을 얻었다. 1) $PGF_{2{\alpha}}$는 $PGE_2$와 같이 적혈구막 삼투성 취약성을 증가시키는데 대조군에서는 NaCl농도 1/18 M 용액에서 완전히 용혈되었으나 $PGE_2$ 및 $PGF_{2{\alpha}}$ 가 $10^{11}M$ 이상 포함될 경우 NaCl농도 $1/16{\sim}1/17\;M$ 에서 100 % 용혈이 일어났다. 2) 동일한 적혈구 부유액을 사용할 때 NaCl농도 1/15 M 용액에서 대조군은 $44.2{\pm}4.3%$가 용혈되나 $PGE_2$ 및 $PGF_{2{\alpha}}$가 $10^{11}M$농도로 포함되었을 때 용혈은 각기 $73.6{\pm}8.4%$ 및 $68.7{\pm}6.4%$로 대조군에 비하여 의의있게 증가하였으며 그 이상의 농도에서는 더 이상 증가를 보이지 않았다. 3) $PGE_2$ 및 $PGF_{2{\alpha}}$에 의해 증가된 용혈은 어느 $Ca^{++}$농도에서도 대조군보다 항상 일정한 정도로 증가되어 있다. 4) 적혈구막 절편에서의 $Ca^{++}$결합은 대조군이나 $PGE_2$ 및 $PGF_{2{\alpha}}$처치군 모두 incubation용액내 $Ca^{++}$농도가 증가함에 따라 곡선적으로 증가하여 $Ca^{++}$농도 5 mM에서 포화된다. 그러나 같은 농도의 $Ca^{++}$에서 비교할 때 적혈구막의 $Ca^{++}$결합은 $PGE_2$ 및 $PGF_{2{\alpha}}$ 존재시 대조군에 비하여 의의있게 증가되었다. 이상의 결과로 보아 $PGE_2$ 및 $PGF_{2{\alpha}}$가 적혈구막의 삼투성 취약성을 증가시키는 기전은 $Ca^{++}$과는 독립적으로 작용함을 알 수 있다.

• KSBB Journal
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• 제27권4호
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• pp.268-272
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• 2012

Microbially induced calcite precipitation (MICP) has been explored for protection and consolidation of construction materials such as concrete. In this study, we isolated 54 calcite forming bacteria from concrete pavement and selected 5 isolates which showed high specific urease activity. Also response of the 5 strains against various environmental stresses was examined. BC 4 and BC 5 showed 35% and 26% viability at heat stress ($50^{\circ}C$), respectively. BC 1 and BC 4 maintained 60.4% and 70.4% viability upon osmotic stress (1 M NaCl), respectively. Among the 5 isolates BC 4 had the highest viability upon alkaline stress (pH 10).

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.92-96
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• 2001

Effect of environmental factors on the expression of soluble forms of Bacillus macerans cyclodextrin glycosyltransferase in recombinant Escherichia coli BL21(DE3)pLysE:pTCGT1 were investigated. The amount of soluble CGTase produced in the cell was measured by determining its enzymatic activity. The soluble fractionof the enzyme was increased by lowering the culture temperature to $30{\circ}C$ and medium pH to 5.8 compared to the enzyme production in LB medium at $37^{\circ}C$ and pH7.0. Addition of 0.2 M NaCl enhanced enzyme expression levels at the expense of cell growth. Glycine betaine that was added after 3 h of induction protected not only the cell growth from hig osmotic pressue but also hepld in vivo folding of CGTase in recombinant E. coli. Addition of 1 mM $CaCl_2$ was also effective in the expression of soluble CGTase, resulting in 15 U/ml of the enzyme activity.

• Journal of Microbiology
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• 제37권3호
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• pp.141-147
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• 1999

Thirteen transcriptionally-fused carbon starvation mutants, derived from Pseudomonas putida ATCC 12633, were analyzed for their survivability and transcriptional induction profiles upon carbon starvation. One of these mutants, MK114, which exhibited the lowest survivability and the highest induction rate, was selected and further examined under different starvation (nitrogen and phosphate) and stress (osmolarity, H2O2, salts, alcohol, and heat) conditions. Under all tested conditions MK114 induced ${\beta}$-galactosidase activity, implying that the interrupted gene (cst114) is a general starvation and stress response gene. The rate of induction ranged from 2.6-fold for phosphate starvation to 3.7-fold for osmotic shock. The mini-Tn5 flanking DNA was cloned from the chromosome of MK114. The cloned DNA fragment exhibited carbon starvation activity, indicating that this fragment contains a carbon starvation-related promoter region. This region was partially sequenced. Possible physiological roles of Cst114 in a carbon sensing mechanism and in other stress responses are also discussed.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1996년도 춘계학술대회
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• pp.286-286
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• 1996

In order to elucidate the mechanism of action of transdermal absorption-enhancing compounds, i.e., pyrrolidone derivatives (2-pyrrolidone, 1-methyl-2-pyrrolidone, 1-ethyl-2-pyrrolidone, 1,5-dimethyl-pyrrolidone and 5-methyl-2-pyrrolidone), multilamellar liposome was prepared from the simulated stratum corneum lipid and employed as a model system for the barrier function of the stratum corneum. The liposomal membrane of the stratum corneum lipid liposome (SCLL) behaves as an osmometer and has an excellent barrier function. In addition, its phase transition temperatures are similar to those of human stratum corneum intercellular lipid region. Therefore, SCLL seems to be a useful skin model. To estimate the barrier function of SCLL, the osmotic behavior of SCLL was measured in the presence of pyrrolidone derivatives and the effect on the phase transition temperature of SCLL was also investigated using differential calorimetry. Above a certain concentration (MLAC), enhances perturb the barrier function of the liposome. The relationship between MLACs and the partition coefficient of the pyrrolidone derivatives was observed; the greater the partition coefficients, the smaller the MLAC. This suggests that the more hydrophobic enhancers penetrate into the lipid layer more easily and reduce the barrier function of membrane more effectively. The results of differential scanning thermograms of the SCLL suggest that the pyrrolidone derivatives had incorporated into the lipid layer in the liposome and increased the fluidity of the lipid layer in the liposome. Such activity might have some correlation with the transdermal absorption-enhancing activity these compounds.

• The Korean Journal of Physiology and Pharmacology
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• 제14권6호
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• pp.377-384
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• 2010

The present study examined whether metformin treatment prevents isoporterenol-induced cardiac hypertrophy in mice. Chronic subcutaneous infusion of isoproterenol (15 mg/kg/24 h) for 1 week using an osmotic minipump induced cardiac hypertrophy measured by the heart-to-body weight ratio and left ventricular posterior wall thickness. Cardiac hypertrophy was accompanied with increased interleukin-6 (IL-6), transforming growth factor (TGF)-${\beta}$, atrial natriuretic peptide (ANP), collagen I and III, and matrix metallopeptidase 2 (MMP-2). Coinfusion of metformin (150 mg/kg/24 h) with isoproterenol partially inhibited cardiac hypertrophy that was followed by reduced IL-6, TGF-${\beta}$, ANP, collagen I and III, and MMP-2. Chronic subcutaneous infusion of metformin did not increase AMP-activated protein kinase (AMPK) activity in heart, although acute intraperitoneal injection of metformin (10 mg/kg) increased AMPK activity. Isoproterenol increased nitrotyrosine levels and mRNA expression of antioxidant enzyme glutathione peroxidase and metformin treatment normalized these changes. These results suggest that metformin inhibits cardiac hypertrophy through attenuating oxidative stress.