• Title/Summary/Keyword: PCR polymorphic

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Morphological characteristics and genetic diversity of Calanthe species native to Korea (한국 자생 새우난초의 형태적 특성 및 유전적 다양성)

  • Cho, Dong-Hoon;Choi, Young-Whan;Kang, Jum-Soon;Lee, Yong-Jae;Choi, In-Soo;Lee, Young-Geun;Jee, Sun-Ok;Kim, Kyung-Min;Son, Beung-Gu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.312-317
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    • 2007
  • This study was conducted to research the morphological characteristics and analyze the genetic diversity by using RAPD in Calanthe species native to Korea. Nine samples were selected by flower color and 19 morphological characteristics. In the length and width of leaf, dorsal sepal, the lateral sepal, the petal, the central lip, and the lateral lip, C. discolor was the shortest and narrowest, respectively, but C. sieboldii was the longest and the widest, respectively. The flower stalk length was the shortest in C. discolor, and the longest in C. sieboldii. Three variants were the intermediate between C. discolor and C. sieboldii in the above morphological characteristics, but spur length was the longest in C. discolor, the shortest in C. sieboldii, and intermediate in the variants. The ovary length in C. discolor was shortest and C. sieboldii and variants were similar with each other. The flower color of C. discolor were brownish red, the value of CIE Lab was between 40 and 50. The flower color of C. sieboldii was yellowish, the value of CIE Lab was between 110 and 130. And variants had various colors between 50 to 70 in the value of CIE Lab. By analyzing multiple band patterns of PCR products, 154 bands were selected as polymorphic RAPD markers. The analysis of genetic similarity of Calanther species using RAPD showed that C. discolor and C. sieboldii are more distant from each other than variants, and these results demonstrated that genetic position of variants located between C. discolor and C. sieboldii.

Genomic Polymorphism Analysis using Microsatellite Markers in Gyeongju Donggyeong Dogs

  • Kim, Seung-Chang;Kim, Lee-Kyung;Choi, Seog-Kyu;Park, Chang-Min;Park, Sun-Ae;Cho, Yong-Min;Lim, Dajeong;Chai, Han-Ha;Lee, Seung-Hwan;Lee, Ji-Woong;Sun, Sang-Soo;Choi, Bong-Hwan
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.243-248
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    • 2012
  • This study was conducted to find a useful marker for gene polymorphism analysis using Microsatellite marker (MS marker) in Gyeongju Donggyeong dog. Twenty three MS marker analyzed the genetic features of DNA using 100 Gyeongju Donggyeong dogs in Gyeongju area. It was performed multiplex PCR with 3 set primer divided 9, 10 and 4 by analysis of conditions among MS markers. The results were calculated heterozygosity, polymorphic information content (PIC), allele frequency and number of allele at each locus using Microsatellite Toolkit software and Cervus 3.0 program. Total 148 alleles were genotyped to determine and average 6.43 alleles was detected. FH3381 had the highest of 15 alleles and FH2834 had the lowest of 2 alleles. Expected heterozygosity had a wide range from 0.282 to 0.876 and had average value of 0.6496. Also, Observed heterozygosity had a more wide range from 0.200 to 0.950 and had average value of 0.6404. PIC had range from 0.262 to 0.859 and average PIC was calculated 0.606. Especially, FH2998 represented the highest rate of observed heterozygosity of 0.950 and FH3381 represented the highest rate of expected heterozygosity of 0.876 and PIC of 0.859. The use of these markers was considered to be useful to study genetic traits of Gyeongju Donggyeong dog.

Polymorphic Variation in Glutathione-S-transferase Genes and Risk of Chronic Myeloid Leukaemia in the Kashmiri Population

  • Bhat, Gulzar;Bhat, Ashaqullah;Wani, Aadil;Sadiq, Nida;Jeelani, Samoon;Kaur, Rajinder;Masood, Akbar;Ganai, Bashir
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.1
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    • pp.69-73
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    • 2012
  • Cancer is a complex disease and the genetic susceptibility to it could be an outcome of the inherited difference in the capacity of xenobiotic metabolizing enzymes. Glutathione-S-transferases (GSTs) are phase II metabolizing enzymes whose various genotypes have been associated with increased risk of different types of cancer. Null mutations caused by the deletion of the entire gene result in the absence of the enzymatic activity and increase in the risk of developing cancer including chronic myeloid leukaemia (CML). In the present case-control study we evaluated the effect of null mutations in GSTM1 and GSTT1 genes on the risk of developing CML. The study included 75 CML patients (43 males and 32 females; age (mean ${\pm}$ S.D) $42.3{\pm}13.4$ years) and unrelated non-malignant controls (76 male and 48 females; age (mean ${\pm}$ S.D) $41.5{\pm}12.9$). The distribution of GSTM1 and GSTT1 genotypes in CML patients and controls was assessed by multiplex-PCR method. Logistic regression was used to assess the relationship between GSTM1 and GSTT1 genotypes and risk of CML. Chi-square test was used to evaluate the trend in modulating the risk to CML by one or more potential high risk genotype. Although GSTM1 null genotype frequency was higher in CML patients (41%) than in the controls (35%), it did not reached a statistical significance (OD = 1.32, 95% CI: 0.73-2.40; P value = 0.4295). The frequency of GSTT1 null genotypes was higher in the CML patients (36%) than in the controls (21%) and the difference was found to be statistically significant (OD = 2.12, 95% CI: 1.12-4.02; P value = 0.0308). This suggests that the presence of GSTT1genotype may have protective role against the CML. We found a statistically significant (OD = 3.09, 95% CI: 1.122-8.528; P value = 0.0472) interaction between the GSTM1 and GSTT1 null genotypes and thus individuals carrying null genotypes of both GSTM1 and GSTT1 genes are at elevated risk of CML.

IL-10 gene promoter polymorphisms in Korean gener-alized aggressive periodontitis patients (한국인 전반적 급진성 치주염 환자에서 IL-10 promoter 유전자 다변성에 관한 연구)

  • Ryn, Ji-Sun;Kim, Ok-Su
    • Journal of Periodontal and Implant Science
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    • v.37 no.3
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    • pp.563-573
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    • 2007
  • Genetic polymorphisms associated with aggressive periodontitis have previously been reported. Interleukin-10 is an immunoregulatory cytokine that plays a role in the pathogenesis of periodontitis. Individual capacity for IL-10 production appears to be under genetic influence, The aim of present investigation was to explore possible genetic association of IL-10 gene promoter polymorphisms with generalized aggressive periodontitis. The study population consisted of 37 generalized aggressive periodontitis patients from the Department of Periodontology, Chonnam National University Hospital and 27 control subjects, all the subjects were non-smokers, Genomic DNA was obtained from buccal swab. The IL-10promoter -597, -824, -1082 positions were genotyped by amplifying the polymorphic regions using polymerase chain reaction (PCR) , followed by restriction enzyme digestion and gel electrophoresis. IL-10-597 C (allele 1) to A (allele 2) and IL-10-824 C (allele 1) to T (allele 2) and IL-10-1082 G (allele 1) to A (allele 2) polymorphisms were examined. The results were as follows. 1. In patients, the distribution of genotypes C/C, C/A and NA at Il-10-597 was determined to be 13.5%, 37.8% and 48.7%, respectively and the distribution of genotypes at IL-10-824 was the same as that of IL-10-597. The distribution of genotypes G/G, G/A and NA at IL-10-1082 was found to be 2.7%, 16.2% and 81. 4%, respectively. No statistical difference in genotype distribution was found between the patient and control groups. 2. Allele 2 carriage rate at the three position of the IL-10 promoter region was higher in the control group than the patient group. 3. Allele 2 frequencies at IL-10-597 and -824 positions were higher in female group than male group and its difference was statistically significant(p<0.05). No significant difference in genotype distribution between the control and patient groups. Allele frequency between control and patient groups was not significantly different although allele 2 frequency at the three positions in the IL-10 promoter region appeared to be higher in control group. In conclusion, no clear association between IL-10 gene promoter polymorphisms and generalized aggressive periodontitis in Korean was observed.

IL-6 gene promoter polymorph isms in Korean generalized aggressive periodontitis (한국인 전반적 급진성 치주염 환자에서 IL-6 유전자 다변성에 관한 연구)

  • Bang, Sun-Jung;Kim, Il-Shin;Kim, Ok-Su;Kim, Young-Jun;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.38 no.4
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    • pp.579-588
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    • 2008
  • Purpose: The purpose of this study was to investigate the association of generalized aggressive periodontitis with IL-6 promoter gene single nucleotide polymorphisms(SNP). Material and Methods: The study population consisted of 52 generalized aggressive periodontitis patients(GAP) and 30 periodontally healthy control subjects, who were systemically healthy non-smokers. Genomic DNA was obtained from buccal swab. The IL-6 promotor SNP at the positions of -597, -572, and -174 were genotyped by amplifying the polymorphic region using polymerase chain reaction(PCR), restriction enzyme digestion and gel electrophoresis. Result: The genotype distributions for G/G, G/A and A/A genotypes of IL-6 -597 were 30.8%, 40.4%, and 28.8% in the GAP group and 53.3%, 40%, and 6.7% in the control group and were statistically different between 2 groups(p<0.05). Allele 2 frequency of IL-6 -597 were significantly higher in the GAP group than the control group(p<0.01). At the position of IL-6 -572, the distribution for C/C, C/G and G/G genotypes were 23.1%, 55.8% and 21.2% in the GAP group and 20%, 33.3%, and 46.7% in the control group. In female subjects, the genotype distribution were significantly different between 2 groups(p<0.01). In male subjects, allele 2 frequency of IL-6-572 was significantly lower in the GAP group than the control group(p<0.05). The genotype distribution of IL-6 -174 in the GAP group were 96.2%, 3.8% for G/G, G/C genotypes whereas only the G/G genotype was detected in the control group. Conclusion: In conclusion, significant associations were found in IL-6 gene promoter(-597, -572) polymorphisms and generalized aggressive periodontitis. Further cohort study will be necessary in larger population.

Construction of a DNA Profile Database for Commercial Cucumber (Cucumis sativus L.) Cultivars Using Microsatellite Marker (Microsatellite 마커를 이용한 오이 유통품종 DNA Profile Data Base 구축)

  • Kwon, Yong-Sham;Choi, Keun-Jin
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.344-351
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    • 2013
  • Microsatellite is one of the most suitable marker for cultivar identification as it has great discrimination power for cultivars with narrow genetic variation. The polymorphism level between 358 microsatellite primer pairs and 11 commercial cucumber cultivars was investigated. Thirty-one primer pairs showed high polymorphism within cucumber cultivars with different fruit types. These markers were applied for the constructing DNA profile data base of 110 commercial cucumber cultivars through multiplex PCR and fluorescence based automatic detection system. A total of 139 polymorphic amplified fragments were obtained by using 31 microsatellite markers. The average of PIC value was 0.610 ranging from 0.253 to 0.873. One hundred and thirty nine microsatellite loci were used to calculate Jaccard's distance coefficients for UPGMA cluster analysis. A clustering group of varieties, based on the results of microsatellite analysis, were categorized into plant shape and fruit type. Almost the cultivars were discriminated by marker genotypes. This information may be useful to compare through genetic relationship analysis between existing variety and candidate varieties in distinctive tests and protection of plant breeders' intellectual property rights through variety identification.

Detection of Pseudomonas tolaasii Causing Brown Blotch Disease of Mushroom with Species-specific DNA Probe (종 특이 DNA probe를 이용한 버섯 세균성 갈반병 병원균(Pseudomonas tolaasii)의 검출)

  • Kwon, Soon-Wo;Go, Seung-Joo;Cheun, Meung-Sook;Kang, Hee-Wan;Oh, Se-Jong;Chang, Who-Bong;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.132-137
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    • 1999
  • This study was carried out to develop the molecular marker for the detection of Pseudomonas tolaasii, a causative agent of bacterial brown blotch disease of oyster mushroom (Pleurotus ostreatus). When several primers designed from repetitive sequences and pectin lyase genes of bacteria were used to produce DNA polymorphism from different Pseudomonas spp. isolated from edible mushrooms, PEU1 primer derived from pectin lyase gene produced polymorphic bands differentiating P. tolaasii strains from other Pseudomonas species. Two bands, 1.0kb and 0.4kb, found commonly in 6 isolates of P. tolaasii were cloned into pGEM-T vector which were designated as pPTOP1 and pPTOP2, respectively, to use as probe. The 0.4 kb insert of pPTOP2 hybridized to only 6 isolates of P. tolaasii, but did not to the other Pseudomonas species. As few as $1.5{\times}10^3$ colony forming unit (cfu) of P. tolaasii could be detected by dot blot hybridization with the cloned 0.4kb DNA in pPTOP2.

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Polymorphism of inter simple sequence repeat markers in Hypsizygus marmoreus (Inter Simple Sequence Repeat(ISSR) 마커를 활용한 느티만가닥버섯(Hypsizigus marmoreus) 종내 다형성 분석)

  • Oh, Youn-Lee;Nam, Yun-Geul;Jang, Kab-Yeul;Kong, Won-Sik;Oh, Min ji;Im, Ji-Hoon;Choi, In-Geol
    • Journal of Mushroom
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    • v.15 no.4
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    • pp.273-278
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    • 2017
  • Hypsizygus marmoreus is a mushroom with abundant flavor and medicinal properties. However, its application is limited by problems such as long cultivation period, low biological efficiency, and microbiological contamination; therefore, there is a substantial need for development of new cultivars of this species. In this study, 55 strains of H. marmoreus were subjected to inter simple sequence repeat (ISSR) analysis to identify markers for the selection of mother strains for breeding from the collected germplasm. ISSR 13 and 15 were confirmed as polymorphic markers. The three strains (KMCC03106, KMCC03107, and KMCC03108) with white cap color were found to be genetically closely related upon UPGMA analysis of both ISSR 13 and 15. Based on the PCR analysis results for ISSR 15, the collected germplasm were differentiated into three groups according to the strain collection year. Thus, ISSR 15 could be a marker for determining the phylogeny of cap color and genetic variations according to the strain collection year. These results suggest that ISSR markers can be effective tools for the selection of mother strains for breeding of H. marmoreus.

Glutathione S-transferase polymorphism of neonatal hyperbilirubinemia in Korean neonates (한국인 신생아 황달과 Glutathione S-transferase 다형성에 관한 연구)

  • Kang, Chang Seok;Hong, Seung Su;Kim, Ji Sook;Kim, Eun Ryoung
    • Clinical and Experimental Pediatrics
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    • v.51 no.3
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    • pp.262-266
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    • 2008
  • Purpose : Glutathione S-transferase (GST) is a polymorphic supergene family of detoxification enzymes that are involved in the metabolism of numerous diseases. Several allelic variants of GSTs show impaired enzyme activity and are suspected to increase the susceptibility to diseases. Bilirubin is bound efficiently by GST members. The most commonly expressed gene in the liver is GSTM1, and GSTT1 is expressed predominantly in the liver and kidneys. To ascertain the relationship between GST and neonatal hyperbilirubinemia, the distribution of the polymorphisms of GSTT1 and GSTM1 were investigated in this study. Methods : Genomic DNA was isolated from 88 patients and 186 healthy controls. The genotypes were analyzed by polymerase chain reaction (PCR). Results : The overall frequency of the GSTM1 null was lower in patients compared to controls (P=0.0187, Odds ratio (OR) =0.52, 95% confidence interval (CI), 0.31-0.88). Also, the GSTT1 null was lower in patients compared to controls (P=0.0014, OR=0.41, 95% CI=0.24-0.70). Moreover, the frequency of the null type of both, in the combination of GSTM1 and GSTT1, was significantly reduced in jaundiced patients (P=0.0008, OR=0.31, 95% CI=0.17-0.61). Conclusion : We hypothesized that GSTM1 and GSTT1 might be associated with neonatal hyperbilirubinemia. However, the GSTT1 and GSTM1 null type was reduced in patients. Therefore the null GSTT1, null GSTM1, and null type of both in the combination of GSTM1 and GSTT1 may be not a risk factor of neonatal jaundice.

Micropropagation and RAPD Analysis of Somaclonal Variants in Lavandula spica cv. Marino (라벤다의 기내증식과 RAPD에 의한 체세포 변이체 분석)

  • Li, Xian Ri;Seong, Eun-Soo;Kim, Il-Seop;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.2
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    • pp.94-100
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    • 1999
  • To establish the mass propagation system of Lavandula spica cv. Marino, shoot tip, node, internode and leaf segment cultures were carried out. RAPD was applied to detect the somaclonal variation. Callus induction was very high in the medium supplemented with 1 mg/l 2.4-D, 2 mg/l NAA. especially and combined with 0.05 mg/l BAP from leaves. Shoot formation was high with $2{\sim}4\;mg/l$ BAP or 4 mg/l BAP + 0.2 mg/l NAA from shoot tip. Shoot proliferation was 9.1 times in the $B_{5}$ medium with 0.5 mg/l BAP and 0.01 mg/l NAA. Root formation was improved in NAA, which was the concentration of 0.1 to 1 mg/l and 1 mg/l IAA. Nursery survival rate was enhanced over 90% and growth was looked good in the acclimation soil consisting of peatmoss : vermiculite : perlite (1:1:1, v:v:v). Randomly amplified polymorphic DNA banding patterns based on polymerase chain reaction (PCR) were used to assess the genetic variation in plants regenerated from in vitro culture.

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