• 대한의생명과학회지
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• 제18권3호
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• pp.310-312
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• 2012

Cytochrome $b_5$ is involved in the reduction of methemoglobin back to hemoglobin, thereby maintaining normal function of the blood to carry oxygen around. Congenital abnormal condition of this enzyme causes a rare disease called methemoglobinemia. At least 4 different retropseudogenes are reported so far for cytochrome $b_5$. However, type III pseudogene has attracted most attention because it contains open reading frame in its structure. Although there is no evidence yet if this pseudogene is actually expressed in the cell or the blood the possibility of its expression needs to be elucidated. We have isolated type III pseudogene by polymerase chain reaction and cloned into pGEX-4T-1 expression vector followed by SDS-PAGE. Protein was expressed and the size of the expressed protein was 28 kDa as expected in its genetic code. This result also shows that the protein is not harmful for the viability of the microorganism. This study may contribute to the genetic diagnosis of cardiac diseases, possibly caused by cytochrome $b_5$.

• Journal of Microbiology and Biotechnology
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• 제22권8호
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• pp.1059-1065
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• 2012

Biocatalytic transfer of oxygen in isolated cytochrome P450 or whole microbial cells is an elegant and efficient way to achieve selective hydroxylation. Cytochrome P450 CYP105P2 was isolated from Streptomyces peucetius that showed a high degree of amino acid identity with hydroxylases. Previously performed homology modeling, and subsequent docking of the model with flavone, displayed a reasonable docked structure. Therefore, in this study, in a pursuit to hydroxylate the flavone ring, CYP105P2 was co-expressed in a two-vector system with putidaredoxin reductase (camA) and putidaredoxin (camB) from Pseudomonas putida for efficient electron transport. HPLC analysis of the isolated product, together with LC-MS analysis, showed a monohydroxylated flavone, which was further established by subsequent ESI/MS-MS. A successful 10.35% yield was achieved with the whole-cell bioconversion reaction in Escherichia coli. We verified that CYP105P2 is a potential bacterial hydroxylase.

• BMB Reports
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• 제40권6호
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• pp.875-880
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• 2007

Glucose oxidase (GOD) is an oxidoreductase catalyzing the reaction of glucose and oxygen to peroxide and gluconolacton (EC 1.1.3.4.). GOD is a widely used enzyme in biotechnology. Therefore the production of monoclonal antibodies and antibody fragments to GOD are of interest in bioanalytics and even tumor therapy. We describe here the generation of a panel of monoclonal antibodies to native and heat inactivated GOD. One of the hybridomas, E13BC8, was used for cloning of a single chain antibody(scFv). This scFv was expressed in Escherichia coli XL1-blue with the help of the vector system pOPE101. The scFv was isolated from the periplasmic fraction and detected by western blotting. It reacts specifically with soluble active GOD but does not recognize denatured GOD adsorbed to the solid phase. The same binding properties were also found for the monoclonal antibody E13BC8.

• International Union of Geodesy and Geophysics Korean Journal of Geophysical Research
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• 제23권1호
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• pp.1-11
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• 1995

Thermospheric neutral winds and temperatures have been collected from the ground-based Fabry-Perot interferometer (FPI) at Thule Air Base ($76.5^{\circ}N{\;}69.0^{\circ}W$), Greenland since 1985. The thermospheric observations are obtained by determining the Doppler characteristics f the [OI] 6300 ${\AA}$ emissions of atomic oxygen. The FPI operates routinely during the winter season, with a limitation in the observation by the existence of clouds. For this study, data acquired from 1985 to 1991 were analyzed. The neutral wind measurements from these long-term measurements are used to investigate the influence of solar cycle variation on the high-latitude thermospheric dynamics. These data provide experimental results of the geomagnetic polar cap are also compared with the predictions of two semiempirical models : the vector spherical harmonics (VSH) model of Killeen et al. (1987) and the horizontal wind model (HWM) of Hedin et al. (1991). The experimental results show a good positive correlation between solar activity and thermospheric wind speed over the geomagnetic polar cap. The calculated correlation coefficient indicates that an increase of 100 in F10.7 index corresponds to an increase in wind speed of about 100 m/s. The model predictions reveal similar trends of wind speed variation as a function of solar activity, with the VSH and HWM models tending to overestimate and underestimate the wind speed, respectively.

• 한국초지조사료학회지
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• 제21권1호
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• pp.21-26
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• 2001

환경 스트레스에 의해 야기되는 활성 산소종에 의한 피해에 내성을 가지는 목초의 개발을 위하여 오차드그래스의 배반 조직 유래의 캘러스에 배추유래의 cytosolic glutathione reductase 유전자(BcGRl)를 Agrobucterium tumefaciens EHA101을 매개로 형질전환시켰다. Hygromcin으로 선발된 캘러스로부터 재분화된 식물체는 야생형과 비교하여 형태적으로 차이를 나타내지 않았다. PCR 및 Southern blot 분석을 통하여 형질전환 식물체의 염색체 내에 BcGRl 유전자가 integration 되었음을 확인하였다. 오차드그래스의 잎으로부터 total RNA를 분리하여 Northern blot 분석을 실시한 결과, 도입된 유전자가 형짙전환 식물체 내에서 지속적으로 발현된다는 것을 확인하였다.

• 한국광물학회지
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• 제15권1호
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• pp.44-58
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• 2002

지금까지 기재되지 않았던 새로운 tridymite의 동질다형상의 하나로 비조화 변조상("IC상")을 확인하였다. IC상의 격자형은 L1상과 같은 대칭성을 가진 단사정계(Bb)로 분석되나, 기하학적인 격자 구조는 오히려 L3샅의 바탕 구조와 같으며 L3상과는 다른 형태의 변조구조를 갖는다(변조 벡터 q=0.22 $c*_{H}$ ; $\lambda$ 37 ).편, Ll상의 전자 회절도형에서 보여주는 독특한 분산 회절곡선은 원자 특히 산소 원자의 동적 비배열(dynamic disorder)이 상온에서도 일어날 수 있음을 암시해 준다. Tridymite 시료의 분쇄에 의한 Ll상에서 L3상으로의 상전이는 점진적이기는 하나 매우 두드러지게 일어난다: Ll$\longrightarrow$LI+IC$\longrightarrow$IC+L3$\longrightarrow$L3. 그러나, 분쇄에 의한 상전이의 구체적인 과정은 각 입자의 국부적인 응력 상태와 미세구조의 발달 상태와 밀접한 관련이 있음이 이번 연구를 통해 드러났다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.818-823
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• 2012

Tall fescue (Festuca arundinacea Schreb.) is an important cool season forage plant that is not well suited to extreme heat, salts, or heavy metals. To develop transgenic tall fescue plants with enhanced tolerance to abiotic stress, we introduced an alfalfa Hsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by polymerase chain reaction, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. The reduced accumulation of thiobarbituric acid reactive substances indicates that the transgenic plants possessed a more efficient reactive oxygen species-scavenging system. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic tall fescue and may be useful in developing stress tolerance in other crops.

• Journal of Microbiology and Biotechnology
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• 제24권4호
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• pp.483-488
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• 2014

In order to improve the expression of heat-resistant xylanase XYNB from Aspergillus niger SCTCC 400264, XynB has been cloned into Pichia pastoris secretary vector pPIC9K. The XynB production of recombinant P. pastoris was four times that of E. coli, and the $V_{max}$ and specific activity of XynB reached $2,547.7{\mu}mol/mg$ and 4,757 U/mg, respectively. XynB still had 74% residual enzyme activity after 30 min of heat treatment at $80^{\circ}C$. From the van der Waals force analysis of XYNB (ACN89393 and AAS67299), there is one more oxygen radical in AAS67299 in their catalytic site, indicating that the local cavity is much more free, and it is more optimal for substrate binding, affinity reaction, and proton transfer, etc, and eventually increasing enzyme activity. The H-bonds analysis of XYNB indicated that there are two more H-bonds in the 33rd Ser of XYNB (AAS67299) than in the 33rd Ala(ACN89393 ), and two H-bonds between Ser70 and Asp67.

• 대한원격탐사학회지
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• 제32권3호
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• pp.221-234
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• 2016

우리나라는 대규모 산업단지와 대도시들이 연안에 집중되면서 연안의 오염이 날로 심각해지고 있다. 이러한 연안 오염을 모니터링하기 위해서 위성 영상을 이용한 연안 수질평가지수 모니터링 연구가 수행될 필요가 있다. 수질평가지수란 저층 산소포화도, 엽록소 농도, 투명도, 용존무기질소 및 용존무기인 농도를 수질평가 항목으로 구성하여 해양환경관리법에 따른 해양환경기준을 통해 해역별로 기준을 설정하여 산출하는 지수이다. 이 연구는 한반도 주변의 연안지역을 대상으로 2011년부터 2013년까지의 현장관측 자료 및 Geostationary Ocean Color Imager (GOCI) 위성 영상을 이용하여 연안 표층 해수에 대한 기계학습 기반의 두 가지 수질평가지수 추정 기법을 개발하였다. 첫 번째 방법으로는 GOCI 반사도를 이용하여 추정된 수질평가 항목들로 수질평가지수를 계산하였고, 두 번째 방법은 GOCI 반사도 및 산출물(엽록소 농도, 총 부유물질, 용존유기물)을 이용하여 수질평가지수를 추정하였다. 기계학습으로는 Random Forest(RF), Support Vector Regression (SVR), Cubist를 사용하였다. 수질평가 항목 추정에서 투명도의 정확도가 가장 높게 나타났으며, 모든 수질평가 항목 추정에서 세 가지 기계학습 중 RF의 정확도가 가장 높았다. 하지만 추정된 수질평가 항목들로 계산한 수질평가지수는 추정된 수질평가 항목들의 오차와 저층 산소포화도의 불확실성으로 인해 정확도가 높지는 않았다. 반면 GOCI 반사도와 산출물을 이용하여 추정한 수질평가지수는 현장 관측 기반 수질평가지수와 비교했을 때 첫 번째 방법보다 정확도가 높게 나타났다. 또한 엽록소 농도가 수질평가지수 추정에 가장 중요한 변수로 나타났다.

• Molecules and Cells
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• 제19권2호
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• pp.191-197
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• 2005

The human immunodeficiency virus type 1 (HIV-1) Tat protein transduction domain (PTD) is responsible for highly efficient protein transduction across plasma membranes. In a previous study, we showed that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. In this study, we fused the human SOD gene with a Tat PTD transduction vector at its N- and/or C-terminus. The fusion proteins (Tat-SOD, SOD-Tat, Tat-SOD-Tat) were purified from Escherichia coli and their ability to enter cells in vitro and in vivo compared by Western blotting and immunohistochemistry. The transduction efficiencies and biological activities of the SOD fusion protein with the Tat PTD at either terminus were equivalent and lower than the fusion protein with the Tat PTD at both termini. The availability of a more efficient SOD fusion protein provides a powerful vehicle for therapy in human diseases related to this anti-oxidant enzyme and to reactive oxygen species.