• Molecules and Cells
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• 제38권4호
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• pp.318-326
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• 2015

We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.

• Clinical and Experimental Reproductive Medicine
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• 제31권1호
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• pp.51-57
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• 2004

Objective: To investigate whether polymorphism of gene encoding COMT is associated with the risk of endometriosis in Korean women. Methods: We investigated 136 patients with histopathologically confirmed endometriosis rAFS stage III/IV and 251 control group women who were surgically proven to have no endometriosis. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of PCR products were done to determine each participant's COMT genotype. Results: The distribution according to NIaIII genetic polymorphisms of COMT were as follows. $COMT^{HH}$, $COMT^{HL}$, and $COMT^{LL}$ genotypes were 56.6% (77 women), 34.6% (47 women) and 8.8% (12 women) in the study group and 50.6% (127 women), 39.4% (99 women) and 10.0% (25 women) in the control group. There was no significant difference between the study group and the control group. Conclusion: The results suggest that COMT genetic polymorphism may not be associated with the development of endometriosis in Korean women.

• 생물정신의학
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• 제11권2호
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• pp.88-93
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• 2004

Background:Serotonin transporter gene-linked polymorphism region(5-HTTLPR) and catechol-O-methyltransferase( COMT) genes are thought to be important factors in some personality traits and the etiology of anxiety disorder. The goal of this study was to determine the role of these genes in personality traits. Method:The participants included 116 healthy adults with no history of psychiatric disorders and other physical illness for the last 6 months. All participants were tested by Temperament and Character Inventory(TCI). The 5-HTTLPR, COMT val158met gene polymorphisms were analyzed with PCR(Polymerase Chain Reaction). Differences on TCI dimensions and sub-scales among groups were examined with t-test and ANOVA. Result:There were possible relationships of the 5-HTTLPR with self-transcendence(P=0.050) and COMT val158met polymorphism with cooperativeness(P=0.053). Conclusion:We found associations between 5-HTTLPR, COMT polymorphisms and the some TCI character dimensions. Further studies of polymorphisms of other genes and their interactions may clarify the complex relationship between personality and genes.

• Journal of the Korean Wood Science and Technology
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• 제38권3호
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• pp.213-222
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• 2010

본 연구에서는 리그닌 전구물질 생합성에 관여하는 CCoAOMT (Caffeoyl-CoA-O-methyltransferase)유전자의 발현을 분자생물학적으로 억제시킨 상태와 정상적인 상태에서 생장한 현사시나무를 대상으로 홀로셀룰로 오스, 리그닌과 단당류 조성에 관한 정량 분석을 각각 실시하였다. 그리고 각각의 목질바이오매스로부터 MWL (milled wood lignin)을 추출하여 메톡실기정량, DFRC, Py-GC/MS, GPC, $^{13}C$ NMR 분석 등 리그닌의 화학 구조적 특성을 비교하였다. 정상재와 형질전환체의 홀로셀룰로오스 함량은 각각 81.6%와 82.3%로 큰 차이는 없었지만, 리그닌 함량은 각각 21.7%와 18.3%로 형질전환체가 약 3.4% 정도 낮았다. 정상재와 형질전환체의 구성 단당류 분석결과, 글루코오스는 각각 511.0 mg/g와 584.8 mg/g, 그리고 자일로오스는 각각 217.8 mg/g와 187.5 mg/g로 나타나 CCoAOMT 발현억제는 단당류 조성에 영향을 미치는 것으로 분석되었다. 각 시 료로부터 단리한 리그닌(MWL)의 메톡실기 정량에 따르면 CCoAOMT 발현을 억제한 시료에서 약 7% 정도의 메톡실기가 감소하는 것으로 나타났고, 이는 $^{13}C$-NMR 분석과 Py-GC/MS 분석을 통하여 확인할 수 있었다. 또한 Py-GC/MS 분석에 의하면 정상재와 형질전환체 리그닌의 G/S 비율이 각각 0.59와 0.44로 나타난점으로 미루어 CCoAOMT 발현 억제는 리그닌 생합성 과정에서 G unit 생합성보다 S unit 생합성에 더 많은 영향을 미치는 것으로 예측할 수 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6591-6596
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• 2014

Background: This meta-analysis was performed to investigate the relationship between methylation of the P14 and O6-methylguanine-DNA methyltransferase (MGMT) genes and the risk of hepatocellular carcinoma (HCC). Materials and Methods: We searched PubMed, EMBASE, the Chinese Biomedical Database (CBM), and the China National Knowledge Infrastructure (CNKI) databases to identify relevant studies that analysed HCC tissues for P14 and MGMT gene methylation status; we then performed a meta-analysis. Odds ratios (ORs) and 95% confidence intervals (95%CIs) were calculated to evaluate the association between gene methylation and the risk of HCC. Results: Ten studies that assessed P14 gene methylation in 630 HCC tumour tissues and nine studies analysing MGMT methylation in 497 HCC tumour tissues met our inclusion criteria. Our meta-analysis revealed that the rate of P14 methylation was significantly higher in HCCs than in adjacent tissues (OR 3.69, 95%CI 1.63-8.35, p=0.002), but there was no significant difference in MGMT methylation between HCC and adjacent tissues (OR 1.76, 95%CI 0.55-5.64, p=0.34). A subgroup analysis according to ethnicity revealed that P14 methylation was closely related to the risk of HCC in Chinese and Western individuals (Chinese, OR 7.74, 95%CI 1.36-44.04, p=0.021; Western, OR 3.60, 95%CI 1.49-8.69, p=0.004). Furthermore, MGMT methylation was not correlated with the risk of HCC in Chinese individuals (OR 2.42, 95%CI 0.76-7.73, p=0.134). The combined rate of P14 methylation was 35% (95%CI 24-48%) in HCC tumour tissues and 11% (95%CI 4-27%) in adjacent tissues, whereas the combined rate of MGMT methylation was 15% (95%CI 6-32%) in HCC and 10% (95%CI 4-22%) in adjacent tissues. Conclusions: These results suggest that the risk of HCC is related to P14 methylation, but not MGMT methylation. Therefore, P14 gene methylation may be a potential biomarker for the diagnosis of HCC.

• 한국초지조사료학회지
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• 제26권1호
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• pp.1-8
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• 2006

조사료로서 소화율을 향상시킨 신품종 형질 전환 오차드그래스를 개발할 목적으로 lignin 합성경로에 있어서 중요한 효소 유전자 중의 하나인 COMT 유전자를 cloning하여 그 특성을 해명하였다. 오차드그래스의 COMT 유전자는 식물체의 전 조직에서 발현되고 있었으며, 특히 줄기와 뿌리조직에서 높은 발현량을 나타냄으로서 목질화에 크게 관여하는 lignin 생합성 유전자일 것으로 판단되었다. Dgcomt 유전자의 발현을 억제시킨 형질전환 오차드그래스를 개발하기 위하여 Dgcomt 유전자를 RNAi 발현벡터에 도입한 후, Agrobacterium 형질전환시스템을 이용하여 오차드그래스에 도입하였다. PCR, Southern 및 Northern 분석 결과 RNAi 발현벡터가 genome에 도입되었으며, Dgcomt 유전자의 발현이 상당한 수준으로 저하되었음을 확인하였다. Dgcomt 유전자의 발현억제는 식물체의 목질화와 더불어 증가되는 lignin의 축적량을 감소시킬 것으로 기대되며 향후 소화율이 증가된 고품질 신품종 목초의 개발에 유용하게 활용될 것으로 판단된다.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.81-81
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• 2003

DNA methylation is a covalent modification of DNA that can modulate gene expression and is now recognized as a major component of the epigenome. During evolution, the dinucleotide CpG has been progressively eliminated from the genome of higher eukaryotes and is present at only 5% to 10% of its predicted frequency. Approxymately 80% of the remaining CpG sites contain methylated cytosines in most vertebrates and they are distributed in a pattern that is unique in each tissue and is inversely correlated with gene expression. The pattern of methylation is faithfully maintained during cell division by the enzyme Dnmt1, the maintenance DNA methyltransferase, which catalyzes the transfer of a methyl group from S-adenosyl-methionine to the 5'-position of the cytosine ring. We have been identified bovine Dnmt1 cDNA full-length recently (AY173048) Little is known on the functions of Dnmt1 in bovine preimplantation embryos. Thus, we analyzed the specific pattern of Dnmt1 in in vitro derived/nuclear transfer bovine and in vivo derived mouse embryos to monitor the epigenetic reprogramming process. We investigated these process by using indirect immunofluresence with an antibody to Dnmt1. According to other studies, Dnmt1 accumulates in nuclei of early growing oocytes but is sequestered in the cytoplasm of mature oocytes. In 2-cell and 4-cell embryos, Dnmt1 is cytoplasmic, but at the 8-cell stage, it is present only in the nucleus. By the blastocyst stage, Dnmt1o is again found only in the cytoplasm. Thus, nuclear localization of Dnmt1o in preimplantation embryos is limited to the 8-cell stages After implantation, Dnmt1 is localized in the nucleus in mouse. However, we have found different patterns of Dnmt1 nuclear localization. Though we used the common antibody, immune-localization data revealed that Dnmt1 antibody have been detected at the nucleus in 1-cell to blastocyst embryos. Therefore, maybe we think that the functions of Dnmt1 between bovine and mice are different. In order to Identify the mechanisms that regulate DNA methylation in bovine preimplantation embryo, we have plans on using bovine oocyte and somatic specific Dnmt1 antibodies.

• Journal of Plant Biotechnology
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• 제36권1호
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• pp.23-29
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• 2009

벼의 저온처리 cDNA 은행에서 분리된 CCCH 형태 zinc finger 단백질인 OsZF2의 기능을 분석하기 위하여 벼에서 CaMV 35S 프로모터 조절하에 OsZF2가 발현(35S:OsZF2)되는 형질전환벼 식물체를 개발하였다. 35S:OsZF2 형질전환벼에 대한 하이그로 마이신저항성 검정을 통해 동형접합체 계통을 선발하고 Northern 발현분석에 의해 OsZF2 유전자가 형질전환체에서 과발현되는 것을 확인하였다. 형질전환체와 대조구인 낙동벼를 100 mM NaCl 첨가 MS 배지에서 키운 후 잎과 뿌리의 길이를 측정하여 내염성 검정을 수행한 결과 대조구에 비해 형질전환체 생육이 다소 양호 한 것으로 나타났다. GMO 포장에서 생육상태를 관찰 한 결과 형질전환체는 생육지연으로 인한 왜화 현상을 나타내며 출수기 또한 열흘 정도 지연되나 등숙기에는 대조구와 같은 초장을 보였다. zinc finger 유전자는 식물체의 발달과 분화 단계 및 환경 스트레스 반응 등 중요한 역할을 하는 것으로 알려져 있으므로 유전체발현 분석으로 하위단계에서 조절되는 유전자 발현 양상을 분석하였다. 35S:OsZF2 전환체에서 낙동벼보다 4배 이상 발현이 증가된 유전자 중에서 게놈 주석에 기초한 기능을 유추하면 신호전달과 관련된 protein kinase, DNA 결합단백질과 대사에 관련된 효소 유전자, 스트레스 반응에 관여하는 일부 유전자 및 병 저항성과 관련된 유전자들의 발현이 증가되었다. 따라서 벼에서 분리된 OsZF2 CCCH type zinc finger 유전자는 벼 성장 발달과 스트레스에 반응하는 상위 조절자로서 기능을 할 것으로 추측된다.

• 동의생리병리학회지
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• 제17권4호
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• pp.905-913
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• 2003

Oriental medicine explains aging as the weakening of Kidney-ai, and Kidney-strengthening herbal medicines such as Yukmijihwang-tang have been studied for anti-aging effects. In Western Medicine, the hypothesis that reactive oxidant species(ROS) contribute to the aging process is generally accepted. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective constituent of Yukmijihwang-tang in decreasing ROS production in oxidative-stressed cells. The purpose of this study is to confirm the anti-oxidant effect of MCR on PC12 cells, the expression of Heme oxygenase (HO), Macrophage migradon inhibitory factor (MIF), Catechol-O-methyltransferase (COMT) using real time RT PCR. PC12 cells were treated without or with hydrogen peroxide in the presence or absence of MCR using MTS assay. Hydrogen peroxide decreased the viability of PC12 cells by 53% and MCR did not influence that of stressed PC 12 cells irrespective of dose or incubation period. However, MCR showed an inhibitory effect on production of ROS in stressed cells, both dose and incubation time dependently. In particular, 1 ㎎/㎖ of MCR for 24 h culture almost returned to normal level. In the quantiation of anti-aging related gene expression, MCR at 1 ㎎/㎖ increased the expression of HO by 370%, MIF by 180% and COMT by 280% through real time RT PCR. In conclusion, MCR treatment protected PC12 cells from hydrogen peroxide and decreased ROS production and enhanced anti-oxidative gene expression such as HO, COMT and MIF, which suggests that MCR is involved in controlling anti-aging of nerve cells through elimination of cytotoxic stimuli.

• 생명과학회지
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• 제27권10호
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• pp.1104-1110
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• 2017

히스톤 H3K4의 메칠화는 3T3-L1의 지방세포의 분화를 촉진하는 것으로 알려져 있으나, 히스톤 H3K4 메칠화 효소인 SET1A가 지방세포 분화를 조절하는지에 대해서는 보고된 바가 없다. 그러므로 본 연구에서는 SET1A의 3T3-L1 지방세포의 분화조절과 기전을 연구하였다. SET1A의 발현은 3T3-L1 지방세포 분화과정에서 증가함을 관찰하였다. 3T3-L1 지방전구세포에서 siRNA을 이용하여 SET1A의 발현을 감소시키면 3T3-L1 지방전구세포의 분화가 억제됨을 관찰하여 SET1A가 3T3-L1 지방전구세포의 분화를 촉진함을 알 수 있었다. 이에 대한 조절기전을 알기 위해, SET1A의 발현을 감소시킨 3T3-L1 지방전구세포의 세포증식을 측정한 결과, 분화 초기 단계인 분화 후 2일 동안 3T3-L1 지방세포의 증식이 감소하였다. 또한 분화 후 7일 동안 지방세포세포 분화 조절인자들의 발현을 측정한 결과, SET1A의 발현을 감소시킨 3T3-L1 지방세포에서 $PPAR{\gamma}$의 발현이 감소하였다. 위와 같은 연구결과를 바탕으로, SET1A는 분화초기단계에서는 mitotic clonal expansion 단계를 촉진하고, 분화후기단계에서는 $PPAR{\gamma}$의 발현을 증가시켜 3T3-L1 지방세포의 분화를 촉진함을 알 수 있었다.