• Journal of Animal Science and Technology
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• 제64권6호
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• pp.1046-1062
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• 2022

This study investigated the effects of L-glutamine (Gln) supplementation on growth performance, physiological traits, heat shock proteins (HSPs), and gene expression related to muscle and adipose tissue development in Hanwoo steers under heat stress (HS) conditions. Eight Hanwoo steers (initial body weight [BW] 570.7 ± 43.6 kg, months of age 22.3 ± 0.88) were randomly separated into two groups, control and treatment, and supplied with the concentration (1.5% of BW kg/day/head) and rice straw (1.5 kg/day/head). The treatment group were fed the Gln supplementation (0.5% of concentration, as-fed basis) once a day at 08:00 h. Blood samples for the assessment of haematological and biochemical parameters and the separation of peripheral blood mononuclear cells (PBMCs) were collected four times, at 0, 3, 6, and 10 weeks of the experiment. Feed intake was measured daily. BW to analyze growth performance and hair follicle collection to analyze the expression of HSPs were executed four times at 0, 3, 6, and 10 weeks. To analyze gene expression, longissimus dorsi muscle samples were collected by biopsy at the end of the study. As a result, growing performance, including final BW, average daily gain, and gain-to-feed ratio, were not different between the two groups. Leukocytes including lymphocytes and granulocytes, tended to increase in the Gln supplementation group (p = 0.058). There were also no differences in biochemical parameters shown between the two groups, except total protein and albumin, both of which were lower in the Gln supplementation group (p < 0.05). Gene expressions related to muscle and adipose tissue development were not different between the two groups. As temperature-humidity index (THI) increased, HSP70 and HSP90 expression in the hair follicle showed a high correlation. HSP90 in the hair follicle was decreased in the treatment group compared with the control group at 10 weeks (p < 0.05). Collectively, dietary Gln supplementation (0.5% of concentration, as-fed basis) may not be influential enough to affect growth performance and gene expression related to muscle and adipose tissue development in steers. However, Gln supplementation increased the number of immune cells and decreased HSP90 in the hair follicle implying HS reduction in the corresponding group.

• 한국환경생태학회지
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• 제30권5호
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• pp.810-820
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• 2016

염생식물 퉁퉁마디의 종자 발아에 영향을 미치는 환경 요인을 조사하고 환경 스트레스에 의해 유도되는 2CysPrx 유전자를 클로닝한 후 스트레스 조건에 따른 2CysPrx 유전자의 발현 양상에 대하여 조사하였다. 염생식물에 대한 가장 대표적인 스트레스는 염분 스트레스로서 퉁퉁마디 발아에 중요한 요인으로 작용하고 있다. 퉁퉁마디의 발아에 대한 NaCl의 한계 농도는 7%로 나타났고, 최적의 발아 조건은 NaCl이 없는 상태로 확인되었다. 퉁퉁마디 발아에서 최적 온도는 $20^{\circ}C$로 98%의 발아율을 보였다. 스트레스에 유도되는 유전자 후보군 중 2CysPrx 유전자의 cDNA를 클론하여 분석한 결과 275개의 아미노산으로 이루어져 있고 두 개의 시스테인 잔기를 가지고 있으며 분자량은 30.1kDa으로 나타났다. 2CysPrx 유전자는 서던 블롯에 의해 유전체에 한 카피 존재하는 것으로 나타났고, 6개의 인트론과 7개의 엑손으로 구성되어 있다. qPCR에 의한 2CysPrx 유전자의 전사율을 분석한 결과, 3.5% NaCl과 40mM $H_2O_2$ 처리 조건에서 전사율이 가장 높게 나타났고, 고온($40^{\circ}C$)과 $75{\mu}M$ ABA 처리 조건에서는 처리 후 8시간에 최고의 전사율을 보였으며, 저온($4^{\circ}C$)에서는 유전자 발현이 일어나지 않는 것으로 나타났다. 우리는 여러 환경 스트레스에 의해 유도되는 다른 유전자의 클로닝을 시도하고 있다.

• Reproductive and Developmental Biology
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• 제29권1호
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• pp.15-18
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• 2005

PSS hetero 돼지를 이용하여 PSS와 관련된 유전자를 cloning 하여 유전자 구조를 분석하고 세포내의 유전자의 존재를 확인하여 PSS 돼지의 유전양식을 밝히고자 실시되었고, 그 결과를 요약하면 다음과 같다. 615번 amino acid가 N과 n에서 각각 arginine과 cysteine으로 존재함을 확인할 수 있었다. 그리고 6번 염색체(PSS 관련유전자)로부터 우성유전자 N과 열성유전자 n이 각각 유전자 좌위(locus)에 대립 유전자(alleles)로 존재함을 확인할 수 있었으며, 이러한 alleles로 존재함으로써 각각의 유전자가 나뉘어져 유전됨을 알 수 있었다.

• 한국어병학회지
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• 제10권1호
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• pp.39-44
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• 1997

Metallothionein은 세균에서 척추동물에 이르기 까지 모든 생명체에 존재하며, 중금속의 세포내농도를 조절하는 중요한 단백질이다. 현재까지 metallothionein의 기능 및 유도기작에 관한 연구는 많이 진척되지는 않았으나, 여러 metallothionein 유전자의 구조가 밝혀져 있는 실정이다. 특히 어류의 metallothionein은 여러종류의 중금속과 환경적인 자극에 의하여 유도되고 정량적인 RT-PCR의 방법으로 metallothionein 유전자의 RNA transcript를 측정함으로써 환경적인 자극의 정도와 중금 속의 상대적인 양을 측정할 수 있기 때문에 중요한 단백질로 인식되고 있다. 본 연구에서는 유전자내부의 특이적 primer와 통상적인 3`말단의 primer를 이용하여 PCR에 의해 450 bp에 해당하는 metallothionein 유전자의 일부의 특성을 조사하였다. 챠넬메기의 cDNA library로부터 PCR에 의해 증폭된 450 bp의 PCR 절편은 다른 어류의 metallothionein 유전자와는 유사성을 보이지 않았다.

• 한국초지조사료학회지
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• 제26권2호
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• pp.83-90
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• 2006

환경스트레스에 내성을 갖는 버즈풋 트레포일(Lotus corniculatus L.) 형질전환체를 개발하기 위하여, AtNDPK 유전자가 E35S 프로모터에 의해 조절되도록 재조합한 발현벡터 pEN-K를 Agrobacterium 형질전환 방법으로 버즈풋 트레포일에 도입하였다. Agrobacterium과 공동배양한 버즈풋 트레포일 캘러스를 $100{\mu}g/m1$의 kanamycin 및 $500{\mu}g/m1$의 cefotaxim을 첨가한 SH-3-kc 배지에서 배양하며 식물체로 재분화시켰다. 재분화된 버즈풋 트레포일의 genomic DNA를 분리, PCR 및 Southern blot 분석을 실시한 결과, AtNDPK 유전자가 도입된 형질전환체의 경우 agarose gel 전기영동 및 X-ray 필름상에서 DNA band 및 hybridization signal을 확인할 수 있었으나, 형질전환되지 않은 대조구의 버즈풋 트레포일에서는 DNA band 및 hybridization signal이 관찰되지 않았다.

• BMB Reports
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• 제42권8호
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• pp.486-492
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• 2009

OsDREB1D, a special DREB (dehydration responsive element binding protein) homologous gene, whose transcripts cannot be detected in rice (Oryza sativa L), either with or without stress treatments, was amplified from the rice genome DNA. The yeast one-hybrid assay revealed that OsDREB1D was able to form a complex with the dehydration responsive element/C-repeat motif. It can also bind with a sequence of LTRE (low temperature responsive element). To analyze the function of OsDREB1D, the gene was transformed and over-expressed in Arabidopsis thaliana cv. Columbia. Results indicated that the over-expression of OsDREB1D conferred cold and high-salt tolerance in transgenic plants, and that transgenic plants were also insensitive to ABA (abscisic acid). From these data, we deduced that this OsDREB1D gene functions similarly as other DREB transcription factors. The expression of OsDREB1D in rice may be controlled by a special mechanism for the redundancy of function.

• 환경생물
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• 제23권1호
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• pp.21-26
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• 2005

Cell response to genotoxic agents is complex and involves the participation of different classes of genes including cell cycle control, DNA repair and apoptosis. In this report, we presented a approach to characterize the cellular functions associated with the altered transcript profiles of MCF-7 exposed to low-dose in vitro gamma-irradiation. We used the method of human 2.4 k cDNA microarrays containing apoptosis, cell cycle, chromatin, repair, stress and chromosome genes to analyze the differential gene expression characterization that were displayed by radiation-exposed cell, human breast carcinoma MCF-7 cell line, such as 4 Gy 4 hr, 8 Gy 4 hr, and 8 Gy 12 hr. Among these genes, 66 were up-regulated and 49 were down-regulated. Specific genes were concomitantly induced in the results. Cyclin dependent kinase 4 (Cdk4) is induced for starting the cell cycle. This regulation is required for a DNA damage­induced G1 arrest. In addition to, an apoptotic pathways gene Bcl-w was concomitantly induced. Mismatch repair protein homologue-l (hMLH1), a necessary component of DNA mismatch protein repair (MMR), in G2-M cell cycle checkpoint arrest. The present study provides new information on the molecular mechanism underlying the cell response to genotoxic stress, with relevance to basic and clinical research.

• 환경생물
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• 제22권3호
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• pp.472-477
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• 2004

본 연구에서는 감마선으로 유도된 돌연변이체들에서 공통으로 발현되는 방사선 관련 유전자들의 발현을 연구하기 위하여, B. lentimorbus WJ5 의 방사선 유도 돌연변이체에서 발현되는 유전자를 DNA microarray로 동시에 탐색하였다. DNA microarray는 B. lentimorbus WJ5 genome을 무작위로 절단하여 2,000 단편으로 구성하였으며, 감마선 $(^{60}/Co)$으로 유도된 7 돌연변이체의 발현을 정량적으로 관찰하였다. 클러스터 분석결과 발현된 408 유전자 중 27개가 감마선 유도 돌연변이체 모두에서 유의하게 발현이 증가되었다. 특히, 복구(mutL, mutM) 에너지 대사 (acsA, sdhB, pgk, yhjB, citB), protease (npr), 산화자극에 대한 환원 (HMM)관련 유전자들이 동시에 증가되었다. 이는 감마선 유도 돌연변이체들에서 자발적인 직/간접 복구 관련 유전자의 발현 증가는 방사선 노출 직후 보이는 stress response와는 다른 현상임을 나타내는 것으로 생각된다.

• Journal of Applied Biological Chemistry
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• 제49권1호
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• pp.1-7
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• 2006

Differential display reverse transcription PCR (DDRT-PCR) analysis was performed on lily 'Marcopolo' bulb scale for isolation of expressed genes during bulblet formation. Cu/Zn lily-superoxide dismutase (LSOD) of 872 bp gene, with ability to scavenge reactive oxygen in stress environment, was isolated. Northern blot analysis showed expression levels of LSOD maximized 12 days after bulblet formation. Ti plasmid vectors were constructed with sense and antisense expressions of LSOD gene and transformed into potato. Southern blot analysis of transgenic potatoes revealed different copies of T-DNA were incorporated into potato genome. In transgenic potatoes, lily SOD gene was overexpressed in sense lines and not in antisense lines. In native polyacrylamide gel electrophoresis analysis, additional engineered LSOD was detected in sense overexpressed transgenic line only. Transgenic potatoes were subjected to oxidative stress, such as herbicide methyl viologen (MV). Transgenic potato lines with sense orientation exhibited increased tolerance to MV, whereas in antisense lines exhibited decreased tolerance. In vitro tuberization of transgenic potato with sense orientation was promoted, but was inhibited in transgenic potato with antisense orientation.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.100-100
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• 2001

Hypoxia is a pathophysiological condition that occurs during injury, ischemia, and stroke. Hypoxic stress induces the expression of genes associated with increased energy flux, including the glucose transporters Glutl and Glut3, several glycolytic enzymes, nitric oxide synthase, erythropoietin and vascular endothelial growth factor. Induction of these genes is mediated by a common basic helix-loop-helix PAS transcription complex, the hypoxia-inducible factor-l${\alpha}$ (HIF-1${\alpha}$)/ aryl hydrocarbon receptor nuclear translocator (ARNT). Insulin plays a central role in regulating metabolic pathways associated with energy storage and utilization. It triggers the conversion of glucose into glycogen and triglycerides and inhibits gluconeogenesis. Insulin also induced hypoxia-induced genes. However the underlying mechanism is unestablished. Here, we study the possibility that transcription factor HIF-1${\alpha}$ is involved in insulin-induced gene expression. We investigate the mechanism that regulates hypoxia-inducible gene expression In response to insulin We demonstrate that insulin increases the transcription of hypoxia- inducible gene. Insulin-induced transcription is not detected in Arnt defective cell lines. Under hypoxic condition, HIF- l${\alpha}$ stabilizes but does not under insulin treatment. Insulin-induced gene expression is inhibited by presence of PI-3 kinase inhibitor and Akt dominant negative mutant, whereas hypoxia-induced gene expression is not. ROS inhibitor differently affects insulin-induced gene expressions and hypoxia-induced gene expressions. Our results demonstrate that insulin also regulates hypoxia-inducible gene expression and this process is dependent on Arnt. However we suggest HIF-l${\alpha}$ is not involved insulin-induced gene expression and insulin- and hypoxia- induces same target genes via different signaling pathway.