• The Plant Pathology Journal
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• 제19권2호
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• pp.123-127
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• 2003

A severe disease of muskmelon (Cucumis melo cv. Alsnight) grown on rockwool in a plastic house was characterized by leaf and stem necrosis followed by death of the plants. In 2001, an isolate of Melon necrotic spot virus-MN (MNSV-MN) of the genus Camovirus was identified as the causal agent of the disease on the basis of biological reactions and nucleotide sequence analyses of coat protein (CP) gene. MNSV-MN induced necrotic local lesions on mechanically inoculated leaves and systemic necrotic spots on the upper leaves of melon cvs. Alsnight, Rui III, Party, Imperial, and Seolhang. However, the inoculated leaves of watermelon and cucumber showed only necrotic lesions. DsRNAs extracted from the melon infected with MNSV-MN were separated into three components. Molecular sizes of the dsRNAs were estimated at approximately 4.5, 1.8, and 1.6 kbp. The amplified cDNA products of CP gene for MNSV-MN by RT-PCR showed approximately 1.2 kbp. The amplified DNA was digested to three fragments by MspI treatment. The cDNA of the genomic RNA of MNSV-MN was cloned and the region deduced to encode the CP was sequenced. The CP coding region, located near 3' end of the genome, consisted of 1,170 nucleotides and had the potential to encode a 390 amino acid protein. The nucleotide and amino acid sequences of MNSV-MN CP gene were 84.0-94.6% and 90.8-94.9% identical with other MNSV isolates found in the GeneBank database, respectively. This is the first report on the occurrence of MNSV in Korea.

• 대한화학회지
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• 제53권6호
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• pp.742-748
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• 2009

정육 사업에서 고기의 원산지와 종을 표기 하는 것은 육질의 판단에 영향을 미친다. 유전자 마커는 종을 판별하기 위한 증거로 사용되므로, 우리는 한우와 젖소 그리고 혼입육을 판단 할 수 있는 유전자 마커의 개발 계획을 수립하였다. 소의 모든 종은 모색 유전자에 의하여 그 종이 결정 되며 모색 유전자의 조절에 의하여 유멜라닌 또는 페오멜라닌이 합성되고, 이로 인하여 모색에 차이가 생기는 점을 이용하여, 종을 판별하는 유전자 마커로 사용된다. 암소와 수소를 판별하기 위하여 Y 염색체 상에 존재하는 성 결정 부위 유전자에 대응하는 프라이머를 제작하였다. 본 연구에서는 모색 유전자와 성 결정 유전자를 다중 중합효소연쇄반응(multiplex-PCR)을 이용하여 증폭하였고, 증폭 산물을 제한 효소 MspA1I로 소화 시켰다. 이 반응물을 변성 고성능 액체 크로마토그래피(denaturing high performance liquid chromatography, DHPLC)를 이용하여 분석하였다. 분석 결과 6가지의 크로마토그램을 확인 할 수 있었고, DHPLC 분석 방법은 한우, 젖소 그리고 혼입육을 쉽게 구별해 낼 수 있었다.

• 농업과학연구
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• 제38권3호
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• pp.453-458
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• 2011

The MC1R (Melanocortin 1 receptor) gene has been known as a causative gene of the coat colors in mammals and responsible for the E (Extension) locus which has three alleles ($E^D$, $E^+$, e) that determines coat colors. The dominant allele $E^D$ produces black or brown colors due to the missense mutation and the recessive e allele has frameshift mutation which shows red or yellow coat colors. Whereas the wild type $E^+$ produces variety of colors due to the interaction with A (Agouti) locus. In this study, PCR-RFLP was performed using two restriction enzymes (BsrF I and MspA1 I) in order to obtain MC1R genotypes in Korean brindle cattle and black cattle. The results showed that all of the animals have the $E^+$ alleles, indicating the $E^+$ allele might related with black coat colors. Later on, the experiments expanded to the 260 Korean candidate bulls whether these animals have the same $E^+$ allele. Among 260 samples investigated, 5% (13/260) of the animals had $E^+$e genotypes, indicating the $E^+$ allele is also present in the candidate bulls in a low frequency. Even though we expected that A locus also affect the black coat color in cattle, all the black coat color animals (brindle and black) have $E^+$ alleles in this study. Therefore, the genotyping of the MC1R gene in candidate bulls will recommended be applied for eliminating of black coat colors in Hanwoo population, if the farmers need to have the brown coat colors only.

• 한국물환경학회지
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• 제24권1호
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• pp.19-29
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• 2008

Hot air sparging is a groundwater remediation technique, in which organic contaminants volatilized into hot air from the saturated to vadose zone. In the laboratory diesel (10,000 mg TPH/kg) was spiked in contaminated saturated aquifer soil. The hot air ($34.9{\pm}2.7^{\circ}C$) was injected in intermittent (Q=1,500 mL/min, 10 minute injection and 10 minute idle) modes. We performed microcosm tests using the groundwater samples to assess TPH reductive remediation activity. For Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis of eubacterial communities in sludge of wastewater treatment plants and soil of experiment site, the 16S rDNA was amplified by Polymerase Chain Reaction (PCR) from the sludge and the soil. The obtained 16S rDNA fragments were digested with Msp I and separated by electrophoresis gel. We found various sequence types for hot air sparging experiment with sludge soil samples that were closely related to Bacillus (149 bp, Firmicutes), Methlobacterium (149 bp, Euryarchaeotes), Pseudomonas (492 bp, ${\gamma}$-Proteobacteria), etc., in the clone library. In this study we find that TPH-water was reduced to 78.9% of the initial value in this experiment aquifer. The results of the present study suggests that T-RFLP method may be applied as a useful tool for the monitoring in the TPH contaminated soil fate of microorganisms in natural microbial community.

• 생물정신의학
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• 제10권2호
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• pp.133-140
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• 2003

Objective:Some candidate gene polymorphisms were reported to be associated with tardive dyskinesia (TD). The aim of this study was to investigate the association of the 5-$HT_{2A}$ receptor gene polymorphisms with TD in Korean schizophrenic subjects. Method:Subjects were of 59 schizophrenic patients with TD and 60 schizophrenic patients without TD for studying of 5-$HT_{2A}$ receptor gene polymorphisms. TD was evaluated using the Abnormal Involuntary Movement Scale(AIMS). Genomic DNA was amplified by PCR and digestion with MspI and BsmI. Result:There were no statistically significant differences in the demographic variables, such as age, male to female percentage, duration of illnesses and duration of antipsychotic drug exposure between the TD group and control group. 1) T102C polymorphisms and TD Comparing the TD group and control group, the 102T/C allele was associated with a significantly increased risk for TD (${\chi}^2$=5.560, df=1, p=0.018). 2) Three AIMS categories of TD and T102C genotype. There were statistically significant differences in the three AIMS categories(${\chi}^2$=6.835, df=2, p=0.033). Conclusion:These result suggest 102T/C genotypes of the 5-$HT_{2A}$ receptor gene are related to the development of TD. The 102T/C genotypes were associated with significantly higher AIMS orofacial dyskinesia scores. These findings suggest that the 5-$HT_{2A}$ receptor gene is significantly associated with susceptibility to TD in patients with chronic schizophrenia.

• 한국환경성돌연변이발암원학회지
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• 제22권4호
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• pp.303-311
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• 2002

This study evaluated the concentrations of urinary metabolites of polycyclic aromatic hydrocarbons (PAHs) in industrial waste incineration workers. The effect of genetic polymorphisms of xenobiotic metabolism enzymes on urinary concentration of PAH metabolites was assessed. And, aromatic DNA adduct levels were also determined in total white blood cells. Fifty employees were recruited from a company handling industrial wastes located in Ansan, Korea: non-exposed group (n=21), exposed group (n=29). Sixteen ambient PAHs were determined by GC/MSD (NIOSH method) from personal breathing zone samples of nine subjects near incinerators. Urinary 1-hydroxypyrene glucuronide (1-OHPG), a major pyrene metabolite, was assayed by synchronous fluorescence spectroscopy after immunoaffinity purification using monoclonal antibody 8E11 (SFS/IAC). Multiplex PCR was used for genotyping for GSTMI/TI and PCR-RFLP for genotyping of CYP1A1 (MspI and Ile/Val). PAH-DNA adducts in peripheral blood WBC were measured by the nuclease P1-enhanced postlabeling assay. Smoking habit, demographic and occupational information were collected by self-administered questionnaire. The range of total ambient PAH levels were 0.00-7.00 mg/㎥ (mean 3.31). Urinary 1-OHPG levels were significantly higher in workers handling industrial wastes than in those with presumed lower exposure to PAHs (p=0.006, by Kruskal-Wallis test). There was a statistically significant dose-response increase in 1-OHPG levels with the number of cigarettes consumed per day (Pearson correlation coefficient=0.686, p<0.001). Urinary 1-OHPG levels in occupationally exposed smoking workers were highest compared with non-occupationally exposed smokers (p=0.053, by Kruskal-Wallis test). Smoking and GSTMI genotype were significant predictors for log-transformed 1-OHPG by multiple regression analysis (overall model R²=0.565, p<0.001), whereas smoking was the only significant predictor for log-transformed aromatic DNA adducts (overall model R²=0.249, p=0.201). Aromatic DNA adducts was also a significantly correlation between log transferred urinary 1-OHPG levels (pearson's correlation coefficient=0.307, p=0.04). However, the partial correlation coefficient adjusting for Age, Sex, and cigarette consumption was not significant (r=0.154, p=0.169). The significant association exists only in individuals with the GSTMI null genotype (pearsons correlation coefficient=0.516, p=0.010; partial correlation coefficient adjusting for age, sex, and cigarette consumption, r=0.363, p=0.038). Our results suggest that the significant increase in urinary 1-OHPG in the exposed workers is due to higher prevalence of smokers among them, and that the association between urinary PAH metabolites and aromatic DNA adducts in workers of industrial waste handling may be modulated by GSTMI genotype. There results remain to be confirmed in future larger studies.

• Asian Pacific Journal of Cancer Prevention
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• 제15권10호
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• pp.4281-4287
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• 2014

Background: Methylation of tumor suppressor genes has been investigated in all kinds of cancer. Tumor specific epigenetic alterations can be used as a molecular markers of malignancy, which can lead to better diagnosis, prognosis and therapy. Therefore, the aim of this study was to evaluate the association between gene hypermethylation and expression of fragile histidine triad (FHIT), glutathione S-transferase P1 (GSTP1) and p16 genes and various clinicopathologic characteristics in primary non-small cell lung carcinomas (NSCLC). Materials and Methods: The study included 28 primary non-small cell lung carcinomas, where an additional 28 tissue samples taken from apparently normal safety margin surrounding the tumors served as controls. Methylation-specific polymerase chain reaction (MSP) was performed to analyze the methylation status of FHIT, GSTP1 and p16 while their mRNA expression levels were measured using a real-time PCR assay with SYBR Green I. Results: The methylation frequencies of the genes tested in NSCLC specimens were 53.6% for FHIT, 25% for GSTP1, and 0% for p16, and the risk of FHIT hypermethylation increased among patients with NSCLC by 2.88, while the risk of GSTP1 hypermethylation increased by 2.33. Hypermethylation of FHIT gene showed a highly significant correlation with pathologic stage (p<0.01) and a significant correlation with smoking habit and FHIT mRNA expression level (p<0.05). In contrast, no correlation was observed between the methylation of GSTP1 or p16 and smoking habit or any other parameter investigated (p>0.05). Conclusions: Results of the present study suggest that methylation of FHIT is a useful biomarker of biologically aggressive disease in patients with NSCLC. FHIT methylation may play a role in lung cancer later metastatic stages while GSTP1 methylation may rather play a role in the early pathogenesis.

• Asian Pacific Journal of Cancer Prevention
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• 제16권2호
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• pp.541-549
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• 2015

Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer (BC) with higher metastatic rate and both local and systemic recurrence compared to non-TNBC. The generation of reactive oxygen species (ROS) secondary to oxidative stress is associated with DNA damage, chromosomal degradation and alterations of both hypermethylation and hypomethylation of DNA. This study concerns differential methylation of promoter regions in specific groups of genes in TNBC and non-TNBC Saudi females in an effort to understand whether epigenetic events might be involved in breast carcinogenesis, and whether they might be used as markers for Saudi BCs. Methylation of glutathione S-transferase P1 (GSTP1), T-cadherin (CDH13), Paired box protein 5 (PAX5), death associated protein kinase (DAPK), twist-related protein (TWIST), DNA-binding protein inhibitor (ID4), High In Normal-1 (HIN-1), cyclin-dependent kinase inhibitor 2A (p16), cyclin D2 and retinoic acid receptor-${\beta}$ ($RAR{\beta}1$) genes was analyzed by methylation specific polymerase chain reaction (MSP) in 200 archival formalin-fixed paraffin embedded BC tissues divided into 3 groups; benign breast tissues (20), TNBC (80) and non-TNBC (100). The relationships between methylation status, and clinical and pathological characteristics of patients and tumors were assessed. Higher frequencies of GSTP1, ID4, TWIST, DAPK, PAX5 and HIN-1 hypermethylation were found in TNBC than in non-TNBC. Hypermethylation of GSTP1, CDH13, ID4, DAPK, HIN-1 and PAX5 increased with tumor grade increasing. Other statistically significant correlations were identified with studied genes. Data from this study suggest that increased hypermethylation of GSTP1, ID4, TWIST, DAPK, PAX5 and HIN-1 genes in TNBC than in non-TNBC can act as useful biomarker for BCs in the Saudi population. The higher frequency of specific hypermethylated genes paralleling tumor grade, size and lymph node involvement suggests contributions to breast cancer initiation and progression.

• Journal of Nutrition and Health
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• 제33권8호
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• pp.813-823
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• 2000

The purpose of this study was to investigate the effects of the nutrition therapy and drug(simvastatin) treatment on the levels of blood lipids for the hyperlipidemic patients according to genetic polymorphism of apo CIII. Subjects of the study consisted of 43(male: 9, female: 34 )hyperlipidemic patients registered to Kyung Hee Medical Center, Intakes of nutrients for the subjects were determined by 24-hr recall method through a personal interviews. The subjects were instructed to take the hypolipidemic and hypocholesterolemic diets. Compliance was monitored through food records, personal interview, and body-weight measurements. The patients changed their basal diet containing 40. 1g fat(182.7mg cholesterol./day) to a diet containing 30.7g fat(139.2mg cholesterol/day) after 12 weeks of nutrition therapy. The distribution of genotypes for apo CIII by Msp I was 32.5% of TT, 50.0% of TC, and 17.5% of CC. aT the beginning of the study, the level of blood triglyceride was the highest for patients with TT and TC. Following the nutrition therapy, patients with CC exhibited the largest drop of blood triglyceride. Following simvastatin treatment with nutrition therapy, blood LDL-cholesterol decreased in most of the patiens with TT. Meanwhile the distribution of genotypes for apo CII by the Fok I was 10.0% of TT, 47.5% of TC, and 42.5% of CC. The level of blood triglyceride was the highest for patients with CC. Following the nutrition therapy, patients with TC exhibited a significant drop for the blood triglyceride. Following simvastatin treatment with nutrition therapy the level of blood LDL-cholesterol decreased in most of the patients with CC. We concluded that dietary habits and food have been changed by nutrition therapy. And blood lipid levels have been decreased by the restriction of intakes of energy, fat and cholesterol. There was variation in the levels of blood lipids according to apo CII polymorphisms. The level of blood lipids responded to nutrition therapy and drug treatment in different was according to genetic polymorphisms. Accordingly, the choice of individualized therapy based on the patient\\`s genetic polymorphism is very important for effective therapy(Korean J Nutrition 33(8) : 813-823, 2000)

• 식물병연구
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• 제14권1호
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• pp.15-20
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• 2008

산박하(Isodon inflexus)의 Is-CMV, 깽깽이풀(Jeffersonia dubia)의 Jd-CMV 및 파리풀(Phryma leptostachya var. asiatica)의 Pla-CMV 등, 3종의 잡초에서 분리한 Cucumber mosaic virus(CMV)를 공시하여, 기주반응 실험, dsRNA 분석, 혈청학적 성질조사, RT-PCR및 RFLP등의 실험을 통하여 각 바이러스를 동정하고, 특성을 구분하였다. 잡초로부터 분리 한 3종의 CMV는 Nicotiana benthamiana, N. tabacum cv. Xanthi nc, N. glutinosa, Cucubita pepo cv. Black Beauty에는 모두 유사한 모자이크 병징을 발현하였으며, Chenopodium amaranticolr와 Vigna unguiculata cv. Kurotanesanzaku에서는 국부 괴사병반이 발현되었다. 한편 고추(Capsicum anmuum cv. Chungyang)에서는 Jd-CMV와 Pla-CMV는 전형적인 모자이크 증상을 발현하였으나, Is-CMV는 병징이 나타나지 않고 무병징으로 감염되는 특성을 보였다. Is-CMV, Jd-CMV 및 Pla-CMV에 감염된 N. benthamiana로부터 추출한 dsRNA는 모두 약 3.4, 3.2, 2.1 및 1.0kbp의 분자크기를 갖는 4종의 dsRNA 밴드가 검출되었으며, 대조로 이용한 Fny-CMV의 dsRNA 패턴과 같았다. Is-CMV, Jd-CMV 및 Pla-CMV에 감염된 N. benthamiana의 즙액을 항원으로 이용하여 Fny-CMV의 항혈청과 한천겔이중확산법으로 조사한 혈청학적 실험 결과는 모든 항원이 한 종의 뚜렷한 침강선을 형성 하였으며, Fny-CMV의 항원에 의해서 형성된 침강선과 융합함으로서 서브그룹 I에 속하는 계통들로 판단되었다. 또한 Is-CMV, Jd-CMV 및 Pla-CMV에 감염된 N. benthamiana로부터 추출한 RNA를 이용하여 CMV-specific 프라이머를 이용한 외피단백질유전자를 포함하는 RNA3의 3' 영역 에 대한 RT-PCR을 실시한 결과, Fny-CMV와 마찬가지로 약 950bp 크기의 cDNA가 증폭되었다. 증폭된 각각의 cDNA를 EcoRI으로 처리하였을 때에는 절단되지 않았으며, HindIII, MspI, SalI 그리고 XhoI에서는 2개의 절편으로 절단되었다. 이와 같은 결과는 Fny-CMV의 절단패턴과 일치하는 것으로 Is-CMV, Jd-CMV 그리고 Pla-CMV는 서브그룹 IA에 속하는 계통으로 확인되었다. 이들 3종의 잡초로부터 CMV가 분리 동정된 것은 이 논문이 처음이다.