• Title/Summary/Keyword: PCR polymorphic

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Polymorphisms of the poly(C)-tract of porcine encephalomyocarditis virus (EMCV) isolated in Korea (국내분리 돼지 뇌심근염바이러스의 poly(C)-tract의 다형태성)

  • Hyun, Bang-Hun;Kim, Hyo-Jin;Kim, In-Joong;Pyo, Hyun-Mi;Kim, Sun-Mi;Kim, Seong-Hee;Kim, Jae-Jo;Lim, Seong-In;Song, Jae-Young
    • Korean Journal of Veterinary Research
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    • v.50 no.3
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    • pp.221-229
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    • 2010
  • Encephalomyocarditis virus (EMCV) belongs to the genus Cardiovirus within the family Picornaviridae. EMCV has been recognized either as a cause of mortality in young pigs, due to acute myocarditis, or of reproductive failure in sows. An EMCV K3 strain was isolated from the heart and brain in a mummified and aborted swine fetus in 1989. For the molecular characterization of the poly(C)-tract of EMCV Korean isolates, K3 strain, viral RNA was extracted and digested with RNase T1, and analyzed the length of the poly(C)-tract by polyacrylamide gel electrophoresis. The poly(C) regions also were amplified by RT-PCR and sequenced. The present study shows that K3 strain of EMCV had a short polymorphic poly(C) tracts (5 to 30 C's) with sequences consisting of $C_9$, $C_{10}$, $C_{13}$, $C_{14}$, $C_{16}$, $C_{20}$, $CUC_{11}$, $C_8UCUC_3UC_{10}$, $C_9UCUC_3UC_{10}$, $C_{10}UCUC_3UC_{10}$, etc. These polymorphism of poly(C)-tracts of EMCV K3 strain implies the historical information of in vivo and/or in vitro passage.

Role of +405C>G and +936C>T Polymorphisms of the Vascular Endothelial Growth Factor Gene and Risk of Esophageal Cancer in the Kashmiri Population

  • Qasim, Iqbal;Bhat, Imtiyaz Ahmad;Masoodi, Khalid Zafar;Shah, Zafar Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.1
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    • pp.97-101
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    • 2015
  • Background: The gene for the vascular endothelial growth factor (VEGF), which promotes angiogenesis and permeability, is polymorphic. The aim of the present study was to evaluate the relationship between +936C>T and +404C>G polymorphism of VEGF with risk of esophageal cancer in the Kashmiri population in India. Materials and Methods: 150 esophageal cancer patients and 150 unrelated healthy controls were genotyped for two VGEF SNPs (+405C/G, and +936C/T) using DNA extracted from prospectively collected blood samples by the PCR-RFLP method. Results: For the VEGF +936C>T polymorphism a significant association of CT and combined CT+TT genotypes was observed with increased risk of esophageal cancer (p=0.021; 0.024). For the +405C>G polymorphism we observed significantly increased frequency of GG genotype in cases as compared to controls and also the +405 GG Genotype was observed to have a two fold risk(OR=2.7356; 95%CI=1.1409-6.5593; p=0.020). The combined genotypes of GG-CC and GG-CT of +405C>G and +936C>T were found to be significantly associated with increased risk of esophageal cancer (p=0.0376; 0.0099). Conclusions: From the results of the present study a significant association of +936C>T and +405C>G polymorphisms with increased esophageal cancer risk exists in the Kashmiri population.

Lack of Influence of MGMT Codon Leu84Phe and Codon Ileu143Val Polymorphisms on Esophageal Cancer Risk in the Kashmir Valley

  • Shah, Mohd A.;Shaffi, Sheikh M.;Lone, Ghulam Nabi;Jan, Syed Mudassar
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3047-3052
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    • 2012
  • The enzyme encoded by the MGMT gene is involved in the repair of alkylated lesions formed in DNA by carcinogenic nitrosamines. Since dietary items consumed by the Kashmiri population contain high concentrations of these agents, it is biologically plausible that MGMT polymorphic variants may be associated with their risk of esophageal cancer. The present study was performed to assess whether non-synonymous SNPS at codon Leu84Phe and codon Ileu143Val of the MGMT gene, close to the active site of the protein, might be linked to predisposition of Kashmiris to esophageal cancer. Genotyping was carried out by polymerase chain reaction-restriction fragment length polymorphism on 92 cases and 77 healthy controls. Codon 84 and codon 143 SNPs of the MGMT gene were not associated with any increase in risk. While the frequency of the Phe allele at codon 84 in cases was (0.16), slightly higher than controls (0.12), the difference was not statistically significant. Similarly, the frequency of Valine allele in cases at codon 143 (0.08) and controls (0.09) was nearly equal. Moreover, no significant association of MGMT genotypes with the clinicopatholgic variables of esophageal cancer patients was observed. In conclusion, MGMT variants at codon 84 and codon143 may not be involved in the susceptibility of the Kashmiri population to esophageal cancer.

Relationships among MTHFR a1298c Gene Polymorphisms and Methylation Status of Dact1 Gene in Transitional Cell Carcinomas

  • Cheng, Huan;Lu, Meng;Mao, Li-Jun;Wang, Jun-Qi;Li, Wang;Wen, Ru-Min;Chen, Jia-Cun
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5069-5074
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    • 2012
  • Objectives: The purpose of this study was to determine the relationship between methylation status of the Dact1 gene and MTHFR a1298c polymorphic forms in transitional cell carcinoma tissues in a Chinese population. Methods: Polymorphisms of folate metabolism enzyme gene MTHFR were assessed by restrictive fragment length polymorphism (RFLP) methods and PCR-based DNA methylation analysis was used to determine the CpG island methylation status of the Dact1 gene. Associations between the methylation status of the Dact1 gene and clinical characteristics, as well as MTHFR a1298c polymorphisms, were analyzed. Results: aberrant methylation of the Dact1 gene was found in 68.3% of cancer tissues and 12.4% of normal tissues,. The methylation rate of the Dact1 gene in cancer tissues was significantly higher in patients with lymph node metastasis than in those without lymph node metastasis (46.3% vs. 17.2%, P = 0.018). No association was found between aberrant DNA methylation and selected factors including sex, age, tobacco smoking, alcohol consumption and green tea consumption. After adjusting for potential confounding variables, variant allele of MTHFR a1298c was found to be associated with methylation of the Dact1 gene. Compared with wild type CC, the odds ratio was 4.33 (95% CI: 1.06-10.59) for AC and 4.95 (95% CI: 1.18-12.74) for AA. The N stage in TNM staging and the occurrence of lymph node metastasis were associated with an MTHFR 1298 AA+AC genotype (P<0.05). Conclusion: MTHFR 1298 AC and AA genotypes might help maintain a normal methylation status of the Dact1 gene, aberrant CpG island methylation of which is closely related to the genesis and progression of transitional cell carcinoma.

Population analysis of eelgrass, Zostera marina L. in Geojedo, Gaedo, and Jedo on the southern coastal water of Korea using RAPD-PCR (RAPD 방법을 이용한 거제도, 개도, 제도해역에서 채집한 말잘피 개체분석)

  • Cho, Eun-Seob;Lee, Sang-Yong;Kim, Jeong-Bae
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.455-461
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    • 2007
  • Assessments of population genetic structure and diversity can be of value in formulating management plans for threatened eelgrass(Zostera maim). Using randomly amplified polymorphic DNA markers, we found evidence of significant genetic structure among the populations of eelgrass sampled at three areas(Geojedo, Gaedo, and Jedo). A highly isolated(>100 km apart) population from the Geojedo had a long genetic distance(0.16), whereas the populations from the Gaedo and Jedo(<10 km apart) exhibited far less distance(0.08). The analysis of similarity within population showed that Geojedo was over 70%, which was of lower value than of Gaedo and Jedo. Based on these results, we realized that heterogeneous population was in accordance with geographic separation. This is caused by limited seed dispersal and interrupted gene flow, although the sample size is small.

Genetic Variation of Korean Lepista nuda (한국산 민자주방망이 버섯의 유전적 변이)

  • 김승희;김종봉
    • Journal of Life Science
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    • v.14 no.1
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    • pp.115-120
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    • 2004
  • Lepista nuda is a world-wide species which has and international reputation as a excellent edible species. In this study, we investigated the genetic variation and taxonomic relationship of L. nuda and other five Tricholomataceae species were analyzed by random amplied polymorphic DNA (RAPD). 15 kinds of random primers were used. The distance matrix was calculated using UPGMA and phyolgenetic relationship were inferred by neighnor-joining (NJ) method. Various bands of 100bp∼1600bp were observed in electrophoretic patterns of RAPD. Nei's genetic distance was calculated using a total of 228 DNA bands identified, and phylogenetic tree was made. The Nei's genetic variations of L. nuda, Lepista surdida, Collybia peronata, Collybia confluens, Lyophyllum cinerascens, Laccara laccata were 0∼21.3%, 21.2∼28.0%, 15.4∼23.0%, 14∼21.8%, 16.5∼34.6%, and 12.4∼27.4%, respectively The consistency index, the retention index and homoplasy index were 0.5217, 0.5769 and 0.5156, respectively. Also, two groups could be made by NJ tree. The genetic distance between L. nuda and C. confluens was closer than that between L. nuda and L. sordida.

Evaluation of Genetic Diversity among Soybean Genotypes Using SSR and SNP

  • Lee, Suk-Ha;P. Tanya;O, Srinives;T. Toojinda;A. Vanavichit;Ha, Bo-Keun;Bae, Jeong-Suk;Moon, Jung-Kyung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.4
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    • pp.334-340
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    • 2001
  • Two different types of molecular markers, simple sequence repeat (SSR) and single nucleotide polymorphism (SNP), were used to measure genetic diversity among five Korean, eight Thai, and three wild soybeans. For SSR analysis, a total of 20 markers were surveyed to detect polymorphisms. For SNP analysis, four primers were designed from consensus sequence regions on disease resistance protein homolog genes, and used to amplify the genomic region. The PCR products were sequenced. A number of polymorphic SSR and SNP bands were scored on all genotypes and their genetic similarity was measured. Clustering analysis was performed independently on both types of markers. Clustering based on SSR markers separated the genotypes into three main groups originated from Korea, Thailand, and wild soybeans. On the other hand, two main groups were classified using SNP analysis. It seemed that SSR was more informative than SNP in this study. This may be due to the fact that SNP was surveyed on the smaller genomic region than SSR. Grouping based on the combined data of both markers revealed similar results to that of SNP rather than that of SSR. This might be due to the fact that more loci from SNP were considered to measure genetic relatedness than those from the SSR.

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Genetic Relationship Based on RAPD Analysis of Yeosu Dolsan Leaf Mustard(Brassica juncea) (RAPD 분석에 의한 여수 돌산갓의 유연관계 분석)

  • Lee, In-Ho;Park, Jong-In;Jeong, Oon-Sup;Jung, Hyo-Jin;Jung, Gun-Ho;Nou, Ill-Sup
    • Journal of Life Science
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    • v.20 no.1
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    • pp.66-70
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    • 2010
  • The genetic relationships of 14 Dolsan leaf mustard (Brassica juncea) lines were analysed using random amplified polymorphic DNA (RAPD) analysis. Thirty nine random primers were tested and seven primers which showed polymorphism were selected. The amplified fragments ranged from 0.2 to 3.5 kb in size. The number of bands amplified in each primer showed the variations ranging from 2 to 27, with an average of 8.7. Based on the UPGMA cluster analysis, 14 lines were separated into three groups. The results showed that RAPD analysis was useful in the genetic diversity and in the cross combination of the $F_1$ hybrid of leaf mustard.

Ethnic Differences in Allelic Frequencies of Two (CA)n Microsatellite Markers Located on Chromosome 5q

  • Hong, Sung-Soo;Chae, Jae-Jin;Goh, Sung-Ho;Yong, Koong-Nam;Lee, Chung-Choo
    • Animal cells and systems
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    • v.1 no.1
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    • pp.123-128
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    • 1997
  • The characteristics of allelic polymorphisms of the two (CA)n microsatellite (p599 and ㅅ599) markers spanning the long arm of chromosome 5 were studied in 52 DNA samples from unrelated inhabitants of Seoul (Korea) by using the polymerase chain reaction (PCR) to investigate differences in allele frequencies between Korean and Caucasian populations. The 6 alleles were observed for p599 (CA)n with a polymorphism informative content (PIC) value of 0.71 and 9 alleles for ㅅ599 (CA)n with a PIC value of 0.82. The observed heterozygote frequencies of the loci were estimated to 0.730 and 0.846, respectively. Several allele frequencies of two loci showed significant differences between Korean and Caucasian populations. Genotype data from the two loci were consistent with the Hardy-Weinberg equilibrium by x2 test. Linkage disequilibrium between p599 (CA)n and ㅅ599 (CA)n loci was observed in x2 test between the observed and expected frequency of allelic association. The probability of matching calculated at each locus was 0.104 for p599 (CA)n and 0.043 for ㅅ599 (CA)n, respectively. These results demonstrate the need to determine populationspecific allele frequency distributions for polymorphic markers when performing genetic linkage studies in racially defined several populations.

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Association between Genetic Polymorphisms of the CYP2C19, CYP2D6 and Types of Sasang Constitutional Medicine (약물대사효소 CYP2C19, CYP2D6의 다형성과 사상체질의 관련성 연구)

  • Lee, Sang-Gyu;Kim, Hyun-Ju;Park, Hye-Jung;Lee, Jung-Ho;Kwon, Deog-Yun;Joo, Jong-Cheon;Choi, Sun-Mi;Lee, Hye-Suk;Kim, Yun-Kyung
    • The Journal of Korean Medicine
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    • v.28 no.1 s.69
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    • pp.51-62
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    • 2007
  • Objectives . The types of Sasang constitutional medicine (SCM) have definite effect on response to herbal drugs. The majority of human P45O dependent xenobiotic metabolism is carried out by polymorphic enzymes which can cause abolished, altered or enhanced metabolism. Therefore, we evaluated the relation of major CYP2C19, 2D6 polymorphism with Sasang types. Methods : 214 healthy subjects were recruited with informed consent; 172 among them had Sasang diagnosis by QSCC2. CYP2D6, 2C19 polymorphism were determined by PCR-RFLP method. Results : None of the Sasang types showed significant difference in CYP2D6, 2C19 polymorphism. However, the Tae-um type showed relatively low frequency of CYP2D6 $^{*}$10/$^{*}$10 polymorphisms with low activity (p=0.110). In the So-yang type, specific $^{*}$3/$^{*}$3 genotype which is a poor metabolizer of CYP2C19$^{*}$3 was detected (p=0.078).Conclusion . These results suggest that the Tae-um type which is said to have high liver function in SCM has the tendency of high drug-metabolizing enzyme activity. With further study, the CYP polymorphism could serve as a scientific tool for SCM diagnosis.

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