• 한국발생생물학회지:발생과생식
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• 제23권3호
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• pp.285-295
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• 2019

Junction-mediating and regulatory protein (JMY) is a regulator of both transcription and actin filament assembly. The actin-regulatory activity of JMY is based on a cluster of three actin-binding Wiskott-Aldrich syndrome protein homology 2 (WH2) domains that nucleate actin filaments directly and promote nucleation of the Arp2/3 complex. In addition to these activities, we examined the activity of JMY generation in early embryo of mice carrying mutations in the JMY gene by CRISPR/Cas9 mediated genome engineering. We demonstrated that JMY protein shuttled expression between the cytoplasm and the nucleus. Knockout of exon 2, CA (central domain and Arp2/3-binding acidic domain) and NLS-2 (nuclear localization signal domain) on the JMY gene by CRISPR/Cas9 system was effective and markedly impeded embryonic development. Additionally, it impaired transcription and zygotic genome activation (ZGA)-related genes. These results suggest that JMY acts as a transcription factor, which is essential for the early embryonic development in mice.

• Parasites, Hosts and Diseases
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• 제58권6호
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• pp.675-679
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• 2020

MYB2 protein was identified as a transcription factor that showed encystation-induced expression in Giardia lamblia. Although nuclear import is essential for the functioning of a transcription factor, an evident nuclear localization signal (NLS) of G. lamblia MYB2 (GlMYB2) has not been defined. Based on putative GlMYB2 NLSs predicted by 2 programs, a series of plasmids expressing hemagglutinin (HA)-tagged GlMYB2 from the promoter of G. lamblia glutamate dehydrogenase were constructed and transfected into Giardia trophozoites. Immunofluorescence assays using anti-HA antibodies indicated that GlMYB2 amino acid sequence #507-#530 was required for the nuclear localization of GlMYB2, and this sequence was named as NLSGlMYB2. We further verified this finding by demonstrating the nuclear location of a protein obtained by the fusion of NLSGlMYB2 and G. lamblia glyceraldehyde 3-phosphate dehydrogenase, a non-nuclear protein. Our data on GlMYB2 will expand our understanding on NLSs functioning in G. lamblia.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.83-83
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• 2003

돼지 난자의 유리화 동결 처리 방법 중 난자를 담는 용기(loading vessel)의 재료로 최근에 알려진 것으로 open-pulled straws(OPS)[Vajta등, Mol Reprod Dev, 51:53-58, (1998)], electron microscope grids(EMG) [Martino등,Biol Reprod, 54:1059-1069, (1996)〕, nylon loop system(NLS) [Lane등, Fertil Steril,72: 1073-1078, (1999)] 등이 보고되고 있다. OPS는 1/4cc straws를 열을 가하여 길게 뽑아 내벽을 얇게 함으로써 filing된 난자나 수정란이 액체 질소와 접촉했을 때 유리화가 신속하게 되도록 하는 방법으로 돼지에서는 별로 보고된 것이 없다. EMG는 열전도가 예민한 전자현미경용 copper grid를 이용한 방법으로 최근 국내 기술진의 연구성적을 포함한 몇몇 학자들에 의하여 보고되었고 NLS는 0.5mm직경의 nylon loop를 이용하여 급속 동결한 성적이 보고되었으나, 돼지 난자에 응용 된 것은 없다. 따라서 이와 같은 동결 재료는 사람과 반추류, mouse외에 돼지 난자에 대하여는 전혀 시도되지 않았지만 유리화 동결기술에서 가장 중요한 실험으로 생각된다. 성공적인 유리화 동결을 위해서는 수정란이 냉각의 전도성이 빠르고, 작은 용액을 수정란과 같이 filling해야 하며 모든 동작이 신속 간편해야 하며 융해 방법도 초급속도의 융해가 요구되므로 이에 부합되어야 한다. 연구 목적은 돼지 난자를 유리화 동결/융해 시 동결 재료-straw/glass, copper grid, nylon 3가지에 대한 제작 방법, 난자 loading, 동결 처리, 보관 방법, 융해 방법 등을 난자의 회수, 수정 후 생존율을 비교 조사하여 가장 우수한 방법을 선택할 목적이었다. 수행 내용은 3가지의 재료의 sample을 제작하고 소독한 다음 준비된 돼지 COCs을 40시간동안 IVM한 후 난자를 5~l5개 정도로 선정 하여 준비된 VS 용액에 평형처리 하였다. 각 재료의 용기에 loading 한 후 동결/보관하였고, 융해는 역순으로 평형하여 maturation 배지에 3~4시간 배양한 다음 경검하고 IVF한 후 NCSU-23 배지에 담아 IVC 배양하면서 cell cleavage상태를 확인하였다.

• Journal of Crop Science and Biotechnology
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• 제11권4호
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• pp.237-242
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• 2008

A liguleless mutant, which showed complete loss of lamina joint region at the junction between leaf blade and leaf sheath, was isolated from a Ds insertional mutants derived from the source cultivar, Dongjin. This mutant could not affect other developmental patterns like phyllotaxis. Southern blot analysis, using GUS as a probe, revealed that the liguleless mutant contained three Ds copies transposed in the rice genome. Among the four genomic sequences flanking the Ds, one was mapped in the intergenic region (31661640 - 31661759), and the other two predicted a protein kinase domain (12098980 - 12098667) as an original insertion site within a starter line used for massive production of Ds insertional mutant lines. Another predicted and inserted in first exon of liguleless 1 protein (OsLG1) that was mapped in coding region (LOC_Os04g56170) of chromosome 4. RT-PCR revealed that the OsLG1 gene was not expressed liguleless mutants. Structure analysis of OsLG1 protein revealed that it predicted transcription factor with a highly conserved SBP domain consisting of 79 amino acids that overlapped a nuclear localization signal (NLS). RT-PCR revealed that OsLG1 is mainly expressed in vegetative organs.

• Bulletin of the Korean Chemical Society
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• 제35권4호
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• pp.1036-1042
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• 2014

In this study, we introduced the RRLR oligopeptide sequences on the surface of polyamidoamine (PAMAM) dendrimer and characterized the physical properties and gene carrier activity of the novel polymer using HEK 293, NIH3T3, and HeLa cells. The RRLR peptide sequences were derived from a mouse fibroblast growth factor 3 (FGF3) protein containing a bipartite NLS motif. The entire sequence of FGF3 is RLRRDAGGRGGVYEHLGGAPRRRK and it has two functional sequences RLRR and RRRK at N-terminus and C-terminus, respectively. In particular, PAMAM G4-RRLR conferred enhanced transfection efficiency and lower cytotoxicity compared with those of PEI 25 kDa, PAMAM G4-R, and PAMAM G4 in various cell lines. These results suggest that the introduction of N-terminal oligopeptides of FGF3 on the surface of PAMAM holds promise as an effective non-viral gene delivery carrier for gene therapy.

• Journal of Plant Biotechnology
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• 제46권2호
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• pp.97-105
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• 2019

Dehydration Responsive Element Binding (DREB) gene is one of the essential transcription factors plants use for responding to stress conditions including salinity, drought, and cold stress. The purpose of this study was to isolate the full length and characterize the DREB gene from three different genotypes of sugarcane, wild, commercial cultivar, and interspecific hybrid sugarcane. The length of the gene, designated ScDREB was 789 bp, and coding for a putative polypeptide of 262 amino acid residues. Sequences of the gene were submitted to the GenBank database with accession numbers of KX280722.1, KX280721.1, and KX280719.1 for wild sugarcane, commercial cultivar (KPS94-13), and interspecific hybrid (Biotec2), respectively. In silico characterization indicated that the deduced polypeptide contains a putative nuclear localization signal (NLS) sequence, and a conserved AP2/ERF domain of the DREB family, at 82-140 amino residues. Based on multiple sequence alignment, sequences of the gene from the three sugarcane genotypes were classified in the DREB2 group. Gene expression analysis indicated, that ScDREB2 expression pattern in tested sugarcane was up-regulated by salt stress. When the plants were under 100 mM NaCl stress, relative expressions of the gene in leaves was higher than those in roots. In contrast, under 200 mM NaCl stress, relative expressions of the gene in roots was higher than those in leaves. This is the first report on cloning the full length and characterization, of ScDREB2 gene of sugarcane. Results indicate that ScDREB2 is highly responsive to salt stress.

• Smart Structures and Systems
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• 제31권1호
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• pp.57-68
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• 2023

Bridge hangers, such as those in suspension and cable-stayed bridges, suffer from cumulative fatigue damage caused by dynamic loads (e.g., cyclic traffic and wind loads) in their service condition. Thus, the identification of damage to hangers is important in preserving the service life of the bridge structure. This study develops a new method for condition assessment of bridge hangers. The tension force of the bridge and the damages in the element level can be identified using the Bayesian optimization method. To improve the number of observed data, the additional mass method is combined the Bayesian optimization method. Numerical studies are presented to verify the accuracy and efficiency of the proposed method. The influence of different acquisition functions, which include expected improvement (EI), probability-of-improvement (PI), lower confidence bound (LCB), and expected improvement per second (EIPC), on the identification of damage to the bridge hanger is studied. Results show that the errors identified by the EI acquisition function are smaller than those identified by the other acquisition functions. The identification of the damage to the bridge hanger with various types of boundary conditions and different levels of measurement noise are also studied. Results show that both the severity of the damage and the tension force can be identified via the proposed method, thereby verifying the robustness of the proposed method. Compared to the genetic algorithm (GA), particle swarm optimization (PSO), and nonlinear least-square method (NLS), the Bayesian optimization (BO) performs best in identifying the structural damage and tension force.

• 생명과학회지
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• 제27권11호
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• pp.1369-1375
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• 2017

본 연구의 목적은 넙치 인지질가수분해효소(PLC-${\delta}1$) 3가지 타입의 세포내 특성을 규명하고자 하였다. 일반적으로 인지질가수분해효소(PLC)의 신호전달경로는 핵, 세포막, 세포질에 분포한다고 알려져 있으나, 핵내 위치 메커니즘은 여전히 불분명하다. PoPLC-${\delta}1A$, PoPLC-${\delta}1B$ (Sf)과 PoPLC-${\delta}1B$ (Lf)의 3타입의 유전자들은 각각 핵위치 신호(NLS)와 핵방출서열(NES)을 포함하고 있다. 본 연구에서는, 넙치 3가지 타입 인지질가수분해효소(PLC-${\delta}1$)의 세포내 위치이동 메커니즘 분석을 위해 GFP 벡터에 유전자를 삽입하여 ionomycin과 thasogargin처리 후 세포위치와 이동양상을 공초점 레이저 주사현미경으로 관찰하였다. PoPLC-${\delta}1A$는 PoPLC-${\delta}1B$ (Lf)와 PoPLC-${\delta}1B$ (Sf)가 원형질막에 국한되어 분포할때 세포질과 세포막보다 세포 소기관에 분포되어 있었다. PoPLC-${\delta}1B$ (Lf) 및 PoPLC-${\delta}1$ (Sf)이 핵 세포질내 이동양상을 보이지 않을 때, PoPLC-${\delta}1A$은 ionomycin과 thapsigargin 처리에 의해 핵 내에 축적되는 양상을 나타냈다. 이런 결과는 손상되지 않은 기능적 NES 서열을 포함하는 PoPLC-${\delta}1A$가 어류에서 핵 세포질 내 왕복 및 이동의 주된 역할을 한다는 것을 보여주고 있다.

• 한국진공학회:학술대회논문집
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• 한국진공학회 1999년도 제17회 학술발표회 논문개요집
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• pp.115-115
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• 1999

GaN는 직접천이형 wide band gap(3.4eV) 반도체로서 청색/자외선 발광소자 및 고출력 전자장비등에의 응용성 때문에 폭넓게 연구되고 있다. 이러한 넓은 분야의 응용을 위해서는 열 적으로 안정된 Ohmic contact을 반드시 실현되어야 한다. n-type GaN의 경우에는 GaN계면에서의 N vacancy가 n-type carrier로 작용하기 때문에 Ti, Al, 같은 금속을 접합하여 nitride를 형성함에 의해서 낮은 접촉저항을 갖는 Ohmic contact을 하기가 쉽다. 그러나 p-type의 경우에는 일 함수가 크고 n-type와 다르게 nitride가 형성되지 않는 금속이 Ohmic contact을 할 가능성이 많다. 시료는 HF(HF:H2O=1:1)에서 10분간 초음파 세척을 한 후 깨끗한 물에 충분히 헹구었다. 그런 후에 고순도 Ar 가스로 건조시켰다. Pd와 Ni은 열적 증착법(thermal evaporation)을 사용하여 p-GaN에 상온에서 증착하였다. 현 연구에서는 열처리에 의한 Pd의 clustering을 줄이기 위해서 wetting이 좋은 Ni을 Pd 증착 전과 후에 삽입하였으며, monchromatic XPS(x-ray photoelectron spectroscopy) 와 SAM(scanning Auger microscopy)을 사용하여 열처리 전과 40$0^{\circ}C$, 52$0^{\circ}C$ 그리고 695$0^{\circ}C$에서 3분간 열처리 후의 온도에 따른 morphology 변화, 계면반응(interfacial reaction) 및 벤드 휨(band bending)을 비교 연구하였다. Nls core level peak를 사용한 band bending에서 Schottky barrier height는 Pd/Ni bi-layer 접합시 2.1eV를, Ni/Pd bi-layer의 경우에 2.01eV를 얻었으며, 이는 Pd와 Ni의 이상적인 Schottky barrier height 값 2.38eV, 2.35eV와 비교해 볼 때 매우 유사한 값임을 알 수 있다. 시료를 후열처리함에 의해 52$0^{\circ}C$까지는 barrier height는 큰 변화가 없으나, $650^{\circ}C$에서 3분 열처리 후에 0.36eV, 0.28eV 만큼 band가 더 ？을 알 수 있었다. Pd/Ni 및 Ni/Pd 접합시 $650^{\circ}C$까지 후 열 처리 과정에서 계면에서 matallic Ga은 온도에 비례하여 많은 양이 형성되어 표면으로 편석(segregation)되어지나, In-situ SAM을 이용한 depth profile을 통해서 Ni/Pd, Pd/Ni는 증착시 uniform하게 성장함을 알 수 있었으며, 후열처리 함에 의해서 점차적으로 morphology 의 변화가 일어나기 시작함을 볼 수 있었다. 이는 $650^{\circ}C$에서 열처리 한후의 ex-situ AFM을 통해서 재확인 할 수 있었다. 이상의 결과로부터 GaN에 Pd를 접합 시 심한 clustering이 형성되어 Ohoic contact에 문제가 있으나 Pd/Ni 혹은 Ni/Pd bi-layer를 사용함에 의해서 clustering의 크기를 줄일 수 있었다. Clustering의 크기는 Ni/Pd bi-layer의 경우가 작았으며, $650^{\circ}C$ 열처리 후에 barrier height는 Pd/Ni bi-layer의 경우에도 Ni의 영향을 받음을 알 수 있었다.

• BMB Reports
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• 제37권5호
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• pp.538-545
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• 2004

A new CRT binding factor (CBF) gene designated Cbcbf25 was cloned from Capsella bursa-pastoris, a wild grass, by the rapid amplification of cDNA ends (RACE). The full-length cDNA of Cbcbf25 was 898 bp with a 669 bp open reading frame (ORF) encoding a putative DRE/CRT (LTRE)-binding protein of 223 amino acids. The predicted CbCBF25 protein contained a potential nuclear localization signal (NLS) in its N-terminal region followed by an AP2 DNA-binding motif and a possible acidic activation domain in the C-terminal region. Bioinformatic analysis revealed that Cbcbf25 has a high level of similarity with other CBF genes like cbf1, cbf2, and cbf3 from Arabidopsis thaliana, and Bncbf5, Bncbf7, Bncbf16, and Bncbf17 from Brassica napus. A cold acclimation assay showed that Cbcbf25 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. Our study implies that Cbcbf25 is an analogue of other CBF genes and may participate in cold-response, by for example, controlling the expression of cold-regulated genes or increasing the freezing tolerance of plants.