• Title/Summary/Keyword: genetic stability

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Characterization of Fusant from Protoplast Fusion between Saccharomyces cerevisiae D-71 and Zygosaccharomyces rouxii SR-S (Saccharomyces cerevisiae D-71과 Zygosaccharomyces rouxii SR-S의 세포융합으로 육성한 융합주의 특성)

  • 이종수;김찬조
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.297-302
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    • 1988
  • The protoplasts of Saccharomyces cerevisiae D-71, a thermophilic strain and Zygosaccharomyces rouxii SR-S, an osmotolerant strain were fused, and a fusant (FS-RN 1) was selected, then was characterized for its genetic stability, DNA content, cell capacity, growth rate, tolerance to salts and chemicals, $\beta$-fructofuranosidase level and ethanol fermenting activity. After 6 months of preservation, 5.8% of the fusant clones were segregated to parental types. The DNA content and cell capacity of the fusant were greater than those of the parental strains. Lag period of growth for the fusant was longer than those for the parents. The fusant colonies showed pink-color reaction to triphenyltetrazolium chloride(TTC) test. The fusant appeared to have resistances to NaCl at moderate levels between both parental strains, and resistances to KCI, sodium propionate and cycloheximide similar to either one of the parents. $\beta$-Fructofuranosidase activity of the fusant was 24.2$\times$10$^{-3}$/IU/g(dry wt) for 3 days culture. Ethanol yields ofter 4 days of fermentation by the fusant at 3$0^{\circ}C$ were : 6.0%(v/v) from 40% of glucose and 8.8%(v/v) from 20% of sucrose.

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Gene Expression in The Fifth Generation of TMV Resistant Transgenic Tobacco Plane at Elevated Temperature (TMV 저항성 형질전환 연초식물체 제 5 세대에서 유전자 안정성 및 고온조건에서의 유전자 발현)

  • 이기원;박성원;이청호;박은경;김상석;최순용
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.245-250
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    • 1998
  • Tobacco mosaic virus(TMV) coat protein cDNA was transformed to Nicotiana tabacum cv. NC82 and the transgenic tobacco plants resistant to TMV infection were isolated in the next generation. The expression of TMV coat protein cDNA and genetic stability of the fifth generation of TMV resistant transgenic tobacco plants at the higher temperature were investigated. The TMV coat protein cDNA was amplified by genomic PCR in all the TMV resistant transgenic tobacco plants. The TMV coat protein expressed in the transgenic tobacco plants was detected at very low level by immunoblot hybridization. Even in tansgenic plants that showed the viral symptom only on very late sucker growth (delay type plants), the coat protein expression in the suckers was much less than that of susceptible tobacco infected with TMV. The TMV coat protein expressed in the transgenic tobacco plants was below 0.01% of total protein. Transcription and expression of the coat protein cDNA in delay type plants were observbed at high temperature (38$^{\circ}C$), and TMV replication was suppressed at both 28$^{\circ}C$ and 38$^{\circ}C$. This indicates that unlike the resistance conferred by 'N' gene. TMV resistance of transgenic tobacco plant won't break down at high temperature.

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DENTAL MANAGEMENT OF A PATIENT WITH MUSCULAR DYSTROPHY UNDER GENERAL ANESTHESIA: CASE REPORT (근이영양증(muscular dystrophy) 환자의 전신마취 하 치과치료 : 증례보고)

  • Chae, Jong Kyun;Song, Ji-Soo;Shin, Teo Jeon;Hyun, Hong-Keun;Kim, Jung-Wook;Jang, Ki-Taeg;Lee, Sang-Hoon;Kim, Young-Jae
    • The Journal of Korea Assosiation for Disability and Oral Health
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    • v.15 no.1
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    • pp.50-54
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    • 2019
  • Muscular dystrophy (MD) is a heterogeneous group of inherited neuromuscular disorders, characterized by progressive muscle weakness. Severity of the disease ranges from mild to severe, and the disease is mostly caused by mutations in a number of genes. These genetic mutations cause lack of proteins which are essential for muscle cell stability. Muscle fibers are gradually replaced by fat and fibrous tissue. The muscles of the head and neck are affected in several types of MD that manifest as altered craniofacial morphology and dental malocclusion. A 3-year-10-month old, 15.0 kg boy with MD presented to Seoul National University Dental Hospital, Seoul, South Korea because of extensive carious teeth. A number of dental caries in primary dentition were identified during clinical oral examination. Due to dental anxiety and underlying systemic disease, general anesthesia was considered. General anesthesia was induced and maintained with intravenous anesthetics, propofol and remifentanil. Caries treatments - resin restoration, pulpectomy, zirconia crown restoration, stainless steel crown restoration - were performed. Under general anesthesia, successful dental procedure was done. Total intravenous anesthesia (TIVA) was performed instead of inhalation anesthesia in order to avoid risk of complications such as malignant hyperthermia and life-threatening rhabdomyolysis. With decreasing muscle function, plaque control becomes more difficult and leads to gingivitis. Especially, the open-mouth posture worsens gingivitis and can leads to malocclusions and problems in swallowing. Regular and periodic dental care is essential for maintaining oral health for patients with MD.

Zearalenone exposure affects the Wnt/β-catenin signaling pathway and related genes of porcine endometrial epithelial cells in vitro

  • Song, Tingting;Yang, Weiren;Huang, Libo;Yang, Zaibin;Jiang, Shuzhen
    • Animal Bioscience
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    • v.34 no.6
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    • pp.993-1005
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    • 2021
  • Objective: Zearalenone (ZEA) has estrogen-like effects. Our previous study has shown that ZEA (0.5 to 1.5 mg/kg) could induce abnormal uterine proliferation through transforming growth factor signaling pathway. To further study the other regulatory networks of uterine hypertrophy caused by ZEA, the potential mechanism of ZEA on porcine endometrial epithelial cells (PECs) was explored by the Illumina Hiseq 2000 sequencing system. Methods: The PECs were treated with ZEA at 0 (ZEA0), 5 (ZEA5), 20 (ZEA20), and 80 (ZEA80) µmol/L for 24 h. The collected cells were subjected to cell cycle, RNA-seq, real-time quantitative polymerase chain reaction, immunofluorescence, and western blot analysis. Results: The proportion of cells in the S and G2 phases decreased (p<0.05), but the proportion of cells in the G1 phase increased (p<0.05) in the ZEA80 treatment. Data analysis revealed that the expression of Wnt pathway-related genes, estrogen-related genes, and mitogen-activated protein kinase pathway-related genes increased (p<0.05), but the expression of genetic stability genes decreased (p<0.05) with increasing ZEA concentrations. The relative mRNA and protein expression of WNT1, β-catenin, glycogen synthase kinase 3β (GSK-3β) were increased (p<0.05) with ZEA increasing, while the relative mRNA and protein expression of cyclin D1 (CCND1) was decreased (p<0.05). Moreover, our immunofluorescence results indicate that β-catenin accumulated around the nucleus from the cell membrane and cytoplasm with increasing ZEA concentrations. Conclusion: In summary, ZEA can activate the Wnt/β-catenin signaling pathway by up-regulating WNT1 and β-catenin expression, to promote the proliferation and development of PECs. At the same time, the up-regulation of GSK-3β and down-regulation of CCND1, as well as the mRNA expression of other pathway related genes indicated that other potential effects of ZEA on the uterine development need further study.

Design Optimization of Multi-element Airfoil Shapes to Minimize Ice Accretion (결빙 증식 최소화를 위한 다중 익형 형상 최적설계)

  • Kang, Min-Je;Lee, Hyeokjin;Jo, Hyeonseung;Myong, Rho-Shin;Lee, Hakjin
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.50 no.7
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    • pp.445-454
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    • 2022
  • Ice accretion on the aircraft components, such as wings, fuselage, and empennage, can occur when the aircraft encounters a cloud zone with high humidity and low temperature. The prevention of ice accretion is important because it causes a decrease in the aerodynamic performance and flight stability, thus leading to fatal safety problems. In this study, a shape design optimization of a multi-element airfoil is performed to minimize the amount of ice accretion on the high-lift device including leading-edge slat, main element, and trailing-edge flap. The design optimization framework proposed in this paper consists of four major parts: air flow, droplet impingement and ice accretion simulations and gradient-free optimization algorithm. Reynolds-averaged Navier-Stokes (RANS) simulation is used to predict the aerodynamic performance and flow field around the multi-element airfoil at the angle of attack 8°. Droplet impingement and ice accretion simulations are conducted using the multi-physics computational analysis tool. The objective function is to minimize the total mass of ice accretion and the design variables are the deflection angle, gap, and overhang of the flap and slat. Kriging surrogate model is used to construct the response surface, providing rapid approximations of time-consuming function evaluation, and genetic algorithm is employed to find the optimal solution. As a result of optimization, the total mass of ice accretion on the optimized multielement airfoil is reduced by about 8% compared to the baseline configuration.

Cis-acting Replication Element Variation of the Foot-and-mouth Disease Virus is Associated with the Determination of Host Susceptibility (구제역바이러스의 숙주 특이성 결정에 연관되어있는 구제역바이러스 cis-acting replication element 변이 분석 연구)

  • Kang, Hyo Rin;Seong, Mi So;Ku, Bok Kyung;Cheong, JaeHun
    • Journal of Life Science
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    • v.30 no.11
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    • pp.947-955
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    • 2020
  • The foot-and-mouth disease virus (FMDV), a member of the Aphthovirus genus in the Picornaviridae family, affects wild and domesticated ruminants and pigs. During replication of the FMDV RNA (ribonucleic acid) genome, FMDV-encoding RNA polymerase 3D acts in a highly location-specific manner. This suggests that specific RNA structures recognized by 3D polymerase within non-coding regions of the FMDV genome assist with binding during replication. One such region is the cis-acting replication element (CRE), which functions as a template for RNA replication. The FMDV CRE adopts a stem-loop conformation with an extended duplex stem, supporting a novel 15-17 nucleotide loop that derives stability from base-stacking interactions, with the exact RNA nucleotide sequence of the CRE producing different RNA secondary structures. Here, we show that CRE sequences of FMDVs isolated in Korea from 2010 to 2017 exhibit A and O genotypes. Interestingly, variations in the RNA secondary structure of the Korean FMDVs are consistent with the phylogenetic relationships between these viruses and reveal the specificity of FMDV infections for particular host species. Therefore, we conclude that each genetic clade of Korean FMDV is characterized by a unique functional CRE and that the evolutionary success of new genetic lineages may be associated with the invention of a novel CRE motif. Therefore, we propose that the specific RNA structure of a CRE is an additional criterion for FMDV classification dependent on the host species. These findings will help correctly analyze CRE sequences and indicate the specificity of host species for future FMDV epidemics.

Molecular Breeding of Tobacco Plants Resistant to TMV and PVY (분자생물학적 TMV 및 PVY 저항성 연초 육종)

  • E.K. Pank;Kim, Y.H.;Kim, S.S.;Park, S.W.;Lee, C.H.;K.H.Paik
    • Proceedings of the Korean Society of Tobacco Science Conference
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    • 1997.10a
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    • pp.134-152
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    • 1997
  • Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

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Variation of Lignan Content for Sesame Seed Across Origin and Growing Environments (참깨 원산지 및 재배지역에 따른 리그난 함량 변이)

  • Kim, Sung-Up;Oh, Ki-Won;Lee, Myoung-Hee;Lee, Byoung-Kyu;Pae, Suk-Bok;Hwang, Chung-Dong;Kim, Myung-Sik;Baek, In-Youl;Lee, Jeong-Dong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.2
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    • pp.151-161
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    • 2014
  • Sesame lignan, including sesamin and sesamolin has been reported to have various content according to accessions and environmental factors. The objective of this study were to analyze the lignan variation of 143 sesame accessions from core collection in Korea and to test the effects of growing years and locations on lignan and lipid content of Korea sesame elite lines. The results showed that the core sesame germplasm in Korea has broad variation of lignan content from 2.33 to 12.17 mg/g with an average 8.18 mg/g. Among tested sesame accessions, the IT184615 had the highest lignan content of as 12.17 mg/g. So this accession will be a good genetic resource for developing a high lignan sesame variety. The sesamin and sesamolin content for sesame accessions across origin had significant difference. The average lignan content of accessions collected from Russia (10.0 mg/g) and Nepal (9.08 mg/g) were relatively higher than other countries. The sesamin and sesamolin content for sesame accessions across seed coat color had significant difference. The average lignan content of sesame with white, brown and black seed coat color was 8.61, 7.51, and 5.49 mg/g, respectively. The variation of lignan and lipid content was significantly different across elite lines, locations and growing years. Therefore, it is important to find sesame accessions having high lignan content with environmental stability.

Characterization of Entomopathogenic Fungus from Trialeurodes vaporariorum and Evaluation as Insecticide (온실가루이 병원성 곰팡이의 특성 및 살충제 개발을 위한 평가)

  • Yoon, Hwi Gun;Shin, Tae Young;Yu, Mi Ra;Lee, Won Woo;Ko, Seung Hyun;Bae, Sung Min;Choi, Jae Bang;Woo, Soo Dong
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.64-70
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    • 2013
  • The greenhouse whitefly, Trialeurodes vaporariorum, is an economically important pest for greenhouse crops because they cause direct damage by feeding on plant nutrients and indirect damage as transmits many virus vectors. It has recently become a serious problem because of the continuous use of insecticide resulting in resistance among greenhouse whitefly population. To overcome these problems, in this study, the biological characteristics and virulence of an entomopathogenic fungus isolated from the cadaver of nymph greenhouse whitefly were investigated. Isolated fungus was identified as Isaria fumosorosea by morphological examinations and genetic identification using sequences of the ITS, ${\beta}$-tubulin, and EF1-${\alpha}$ regions. This fungus was named as I. fumosorosea SDTv and tested for the virulence against nymphs T. vaporariorum and the cold activity, the thermotolerance and the stability of UV-B irradiation on conidia. Mortality rate of greenhouse whitefly showed from 84 to 100% and the virulence increased with increasing conidial concentrations, $1{\times}10^5$ to $10^8$ conidia/ml. Conidia were stable at $35^{\circ}C$, 0.1 $J/cm^2$ of UV irradiation and germinated after 8 days at $4^{\circ}C$. Additionally, the activities of chitinases and proteases produced by I. fumosorosea SDTv were varied according to the medium. In conclusion, I. fumosorosea SDTv which showed high mortality rate against greenhouse whitefly will be used effectively in the integrated pest management programs against the greenhouse whitefly.

Biochemical Characterizations of Phenylalanine Ammonia-Lyase and its Mutants to Develop an Enzymatic Therapy for Phenylketonuria (페닐케톤뇨증의 효소치료 개발을 위한 phenylalanine ammonia-lyase 및 유전자 변이형의 생화학적 특성)

  • Kim, Woo-Mi
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1226-1231
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    • 2009
  • Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.