• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1266-1275
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• 2010

Isozyme and protein electrophoresis data from mycelial extracts of 27 isolates of Trichoderma harzianum, 10 isolates of T. aureoviride, and 10 isolates of T. longibrachiatum from Southern Peninsular Malaysia were investigated. The eight enzyme and a single protein pattern systems were analyzed. Three isozyme and total protein patterns were shown to be useful for the detection of three Trichoderma species. The isozyme and protein data were analyzed using the Nei and Li Dice similarity coefficient for pairwise comparison between individual isolates, species isolate group, and for generating a distance matrix. The UPGMA cluster analysis showed a higher degree of relationship between T. harzianum and T. aureoviride than to T. longibrachiatum. These results suggested that the T. harzianum isolates had high levels of genetic variation compared with the other isolates of Trichoderma species.

• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1557-1563
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• 2005

Ovarian follicular fluid contains both stimulatory and inhibitory agents that influence the growth and maturation of oocyte. In the present study, an attempt was made to isolate and study the biological properties of ovarian follicular fluid peptide(s) in buffaloes. Bubaline ovarian follicular was made steroid- and cell-free. A protein fraction was obtained by saturation (30-35% level) of the follicular fluid with ammonium sulfate. The protein fraction was purified with Sephadex-G 50 gel filtration chromatography and a single peak was obtained in the eluant volume, which was lyophilized. SDS-PAGE of the lyophilized fraction revealed a single band and the molecular weight of the peptide was 26.6 kDa. The peptide stimulated the cumulus cell expansion and in vitro maturation rate of oocytes in buffaloes in a dose dependent manner when it was incorporated at different dose levels (0, 10, 25, 50, 100 and 1,000 ng $ml^{-1}$ of maturation medium). The basic culture medium consisted of TCM 199 with Bovine serum albumin (0.3%). The in vitro maturation rates were comparable to those obtained with a positive control medium (TCM 199+20 ng EGF $ml^{-1}$+steer serum (20%)). Further purification and biological assays may throw more light on the nature and functions of this peptide.

• 한국미생물·생명공학회지
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• 제7권4호
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• pp.211-216
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• 1979

단세포 단백질의 합성이나 환성오니법에 의한 폐수처리 등에 응용되고 있는 Air-lift Fermentor (ALF)는 일반적인 기계식 교반 발효조에 비하여 산소전달 효과가 우수한 것으로 알려져 있다. 그러나 미생물의 성장반응에는 적절한 교반이 필요하므로 발효조 내에서는 높은 산소 농도와 함께 활발한 교반이 이루어져야 한다. 본 연구에서는 ALF 내에서의 산소전달에만 국한해오던 연구 방향에서 실제로 Methylomonas, sp를 이용한 methanol fermentation을 행하여 이 발효조의 기능을 조사하였다. 그 결과 교반은 활발히 일어나고 있으나 높은 농도의 산소를 얻기 위해서는 높은 압력하에서나 낮은 dilution rate에서 운전하여야 하는 것으로 나타났다.

• Journal of Animal Science and Technology
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• 제48권5호
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• pp.719-726
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• 2006

본 실험은 가수분해 시킨 유청단백질과 Lb. casei HY2782의 세포 추출물을 인간 전립선 세포에처리하여 세포 내 glutathione 농도 상승효과와 산화제 tbutyl hydroperoxide (TBHP)에 의한핵산 손상 방지 효과를 알아보기 위한 실험으로 전립선 세포 RWPE1 cells와 PC3MMM2 cells에 hydrolyzed WPI(500g/m)를 48시간 동안 처리 시, 가수분해 시키지 않은 WPI 보다 각각 28.2%, 38.4%씩 GSH 농도가 증가하는 것을 유의적으로 확인할 수 있었고(p<0.05), 카제인의 경우 RWPE1 cells and PC3MMM2 cells에 처리 시 가수분해 시킨 카제인과의 유의차가 보이지 않았으며 (p<0.05), glutathione 합성저해제인 Buthionine sulfoximin (BSO) 처리 시 Glutathione 농도가 현저하게 낮아지는 것을 확인할 수가 있었다(Anderson, 1998). 전립선 세포 RWPE1 cells 와 PC3MMM2 cells에 Lb. casei HY2782 세포 추출물 처리 시 PC3MMM2 cell에서는 GSH의 농도가 유의적으로 증가하는 것을 확인 할 수 있었고(p<0.05), RWPE1 cell에서는 증가 성향을 확인 할 수가 있었다.가수분해 시킨 유청단백질과 Lb. casei cell 세포추출물 처리 시 산화제에 의한 세포의 DNA 손상 억제정도와 사멸율의 변화를 확인 한 결과, 가수분해 WPI를 처리 후 Oxidant tbutyl hydroperoxide (TBHP)(500mM)를 이용하여 PC3MMM2 세포에 산화적인 스트레스를 주었을 시, TBHP만 처리한 구에서는 62.0%의 생존율을 나타내었고, glutathione 합성저해제인 BSO를 첨가시킨 처리구에서는 33.7%의 생존율을 나타내었다. 가수분해 시킨 WPI와 카제인 처리구에서는 각각 86.7%, 63.6%의 생존율을 나타내었고, WPI와 BSO를 함께 처리한 구에서는 71.5%의 생존율을 나타내었다. 이는 BSO에 의해 GSH의 생성이 저해되어 나타난 결과로 보여지며(Anderson, 1988), 가수분해시킨 WPI 가 가수분해 시킨 카제인보다는 높은 생존율을 나타내는 것을 유의적으로 확인할 수가 있었다. (p<0.05) Lb. casei 세포 추출물처리에 의한 생존율에서는 가수분해시킨 WPI와 유사한 수준의 82.4%의 생존율을 보였다. 가수분해시킨 유청단백질과 Lb. casei cell 세포추출물에 의한 세포에서의 glutathione 농도 증가에 의해 산화제에 의한 DNA의 손상이 적었던 것으로 사료되어지고, 이로 인해 생존율이 높았던 것으로 판단된다.

• BMB Reports
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• 제32권1호
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• pp.92-97
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• 1999

Cytokinesis and septation should be coordinated to nuclear division in the cell division cycle for precise transmission of the genome into daughter cells. byr4, an essential gene in fission yeast Schizosaccharomyces pombe, regulates the timing of cytokinesis and septation in a dosage-dependent manner. We examined the intracellular localization of the Byr4 protein by expressing byr4 as a fusion of green fluorescence protein (GFP). The Byr4 protein localizes as a single dot on the nuclear periphery of interphase cells, duplicates before mitosis, and the duplicated dots segregate with the nuclei in anaphase. The behavior of Byr4p throughout the cell cycle strongly suggests that Byr4p is localized to the spindle pole body (SPB), a microtubule organizing center (MTOC) in yeast. The presence of the Byr4 protein in the SPB is consistent with its function to coordinate mitosis and cytokinesis. We also mapped the domains of Byr4p for its proper localization to SPB by expressing various byr4 deletion mutants as GFP fusions. Analyses of the diverse byr4 deletion mutants suggest that the indirect repeats and the regions homologous to the open reading frame (ORF) YJR053W of S. cerevisiae in its C-terminus are essential for its localization to the SPB.

• Molecules and Cells
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• 제38권1호
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• pp.33-39
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• 2015

The correction of disease-causing mutations by single-strand oligonucleotide-templated DNA repair (ssOR) is an attractive approach to gene therapy, but major improvements in ssOR efficiency and consistency are needed. The mechanism of ssOR is poorly understood but may involve annealing of oligonucleotides to transiently exposed single-stranded regions in the target duplex. In bacteria and yeast it has been shown that ssOR is promoted by expression of $Red{\beta}$, a single-strand DNA annealing protein from bacteriophage lambda. Here we show that $Red{\beta}$ expression is well tolerated in a human cell line where it consistently promotes ssOR. By use of short interfering RNA, we also show that ssOR is stimulated by the transient depletion of the endogenous DNA mismatch repair protein MSH2. Furthermore, we find that the effects of $Red{\beta}$ expression and MSH2 depletion on ssOR can be combined with a degree of cooperativity. These results suggest that oligonucleotide annealing and mismatch recognition are distinct but interdependent events in ssOR that can be usefully modulated in gene correction strategies.

• 한국식품영양학회지
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• 제30권6호
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• pp.1279-1285
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• 2017

$Amyloid-{\beta}$ protein ($A{\beta}$) is known to increase free radical production in neuronal cells, leading to cell death by oxidative stress. The purpose of this study was to evaluate the protective effects of $PineXol^{(R)}$ on $A{\beta}_{25-35}$ induced neuronal cell death. Rat pheochromocytoma (PC-12) cells were pre-treated with $100{\mu}g/mL$ of $PineXol^{(R)}$ for 2 h. The cells were exposed to single dose of $30{\mu}M$ $A{\beta}_{25-35}$ for 24 h. Cell death was assessed by a cell count kit-8 (CCK-8) assay, lactate and dehydrogenase (LDH) release assay. An Apoptotic process was analyzed by a protein expression of the Bcl-2 family using western blotting. Cell viability increased in PC-12 cells treated with both $A{\beta}_{25-35}$ and $PineXol^{(R)}$, compared to the control group. $PineXol^{(R)}$ induced a decrease of the Bcl-2 protein expression (p<0.05), while Bax and Sod1 increased (p<0.05), indicating attenuation of $A{\beta}_{25-35}$ induced apoptosis. These results suggest that $PineXol^{(R)}$ may be a good candidate for the prevention of Alzheimer's disease(AD).

• International Journal of Industrial Entomology and Biomaterials
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• 제24권1호
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• pp.23-27
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• 2012

Porcine circovirus type 2 (PCV2) is a single-stranded circular DNA virus associated with Postweaning multisystemic wasting syndrome (PMWS), which is considered to be an important infectious swine viral disease. PCV2 capsid protein encoded by ORF2 is a structural protein and expected as the high immunogenicity protein. In this study, we generated recombinant baculovirus containing ORF2 of PCV2 and analyzed the optimal conditions for the production of capsid protein in insect cell. Production and status of recombinant capsid protein in insect cell were confirmed by SDS-PAGE and Western blot analysis using His tag antibody and anti-PCV2 serum. The yield of recombinant capsid protein was high like as shown visible on SDS-PAGE. Optimal multiplicity of infection (MOI) and infection time of recombinant virus were determined as 5 MOI and 4 days, respectively. ORF2 is known to have N-linked glycosylation site, but we couldn't detect the glycosylation of recombinant protein in insect cells.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.77.1-77
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• 2003

We have isolated a full-length cDNA clone, CaCDPK4 encoding a typical calcium-dependent protein kinase (CDPK) from hot pepper cDNA library. Genomic southern blot analysis showed that it belongs to a multigene family, but represents a single copy gone in hot pepper genome. RNA expression pattern of this gene revealed that it is induced by infiltration of Xanthomonas axonopodis pv. glycines Bra into hot pepper leaves but not by water deficit stress. However, high salt treatment of NaCl (0.4 M) solution to hot pepper plants strongly induced CaCDPK4 gene. In addition, this gene is weakly responsive to the exogenous application of salicylic acid or ethephon. Biochemical study of the GST-CaCDPK4 recominant protein showed that it autophosphorylates in vitro and the presence of EGTA, a calcium chelater, eliminates the kinase activity of the recombinant protein. As a way to identify the in vivo function of CaCDPK4 in plants, VIGS (Virus-Induced Gene Silencing) was employed. Agrobacterium-mediated TRV silencing construct containing the kinase and calmodulin domain of CaCDPK4 resulted in cell death of Nicotiana benthamiana plants. A highly homologous H benthamiana CDPK gene, NbCDPK5, to CaCDPK4 was cloned from N. benthamiana cDNA library. VIGS of NbCDPK5 also resulted in cell death. The molecular characterization of this cell death phenotype is being under investigation.

• Applied Biological Chemistry
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• 제48권4호
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• pp.301-310
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• 2005

구제역(Foot-and-Mouth Disease: FMD)이란 소, 돼지, 양, 염소 등의 cloven-hoofed 동물에서 나타나는 바이러스성 질병으로 입, 코, 유두, 발굽 등에 수포가 형성되는 것이 특징이다. 일곱 가지 혈청형(O, A, C, Asia1, SAT1, SAT2 and SAT3)으로 분류되는 구제역바이러스(Foot-and-Mouth Disease Virus: FMDV)는 single stranded positive RNA virus로 nonenveloped capsid virus이다. Viral genome은 8.2 Kb로 하나의 ORF인 polyprotein으로 되어있으며, 크게 capsid protein coding region인 P1, replication related protein coding region인 P2, RNA dependent RNA polymerase coding region인 P3로 구성된다. FMDV는 respiratory tract의 pharynx epithelial cell에 감염되며, lung epithelial cell에서 replication을 한다. 구제역바이러스는 감염율은 높지만 낮은 치사율을 가진다. 2002년 한국에서 구제역이 발병하여 많은 경제적 손실을 입었다. FMDV의 감염을 조절할 수 있는 조절방법이 없는 실정이며, 현재 많은 나라에서는 구제역바이러스의 감염을 막을 수 있는 효과적인 방법을 연구하고 있다. 본 보고서에서는 FMD에 대한 보다 효과적인 예방법인 DNA vaccine, edible vaccine, peptide vaccine에 대해 고찰하였다.