• Title/Summary/Keyword: DNA restriction

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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.

Comparison of Biological Characteristics on the Organic Waste-treated Lysimeter Soil by RFLP, PLFA, and CLSU (RFLP, PLFA, CLSU를 이용한 폐기물연용토양의 토양미생물 특성 평가 비교)

  • Jang, Kab-Yeul;Weon, Hang-Yeon;Lee, Kang-Hyo;Kwon, Sun-Ik;Kong, Won-sik;Suh, Jang-sun;Sung, Jae-Mo
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.6
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    • pp.415-418
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    • 2008
  • The application of sludge wastes into agricultural fields has been increasing annually in Korea. In particular, sewage sludge application has been widely accepted in decades. Sewage sludge application aid in the recycling of essential nutrients and act as a source of organic matter improving the structure and water-holding properties of the soil. The efficient use of sludge wastes, however, requires an individual assessment of waste products. This study assessed the biological characteristics of organic waste-treated lysimeter soils and develop its indicator to assess the soil health of organic waste-treated lysimeter soils. Several analytical techniques more recently developed such as restriction fragment length polymorphism (RFLP), phospholipid fatty acid (PLFA), and community level substrate utilization (CLSU) fingerprints allow for detailed analyses of soil microbial communities. PLFA and RFLP was, therefore, used in the study to characterize the microbial communities in soil without the need to isolate individual fungi and bacteria. PLFA, RFLP and CLSU have been utilized to assess microbial characteristics of the lysimeter soils with four different sludge wastes for eight consecutive years. Each of these methods was analyzed for a different aspect of soil microbial characteristics. The study would disclose those methods yielded highly reproductive results for each soil and allow distinguishing the soils based on the structures of specific geneand PLFA-pools more than CLSU fingerprints. PLFA methods, especially, revealed the same relative similarities of the treated soils based on cluster analysis of the biological characteristics. Pig manure compost-treated soil, however, was only the same relative resemblance among the three methods. These results indicated that PLFA easily assessed the biological soil characterization.

Analysis of the Genotype Distribution in Cattle Breeds Using a Double Mismatched Primer Set that Discriminates the MC1R Dominant Black Allele (소 MC1R 우성흑모색 대립인자를 구분하는 변형 프라이머를 이용한 소 품종들의 유전자형 분포 분석)

  • Han, Sang-Hyun;Kim, Young-Hoon;Cho, In-Cheol;Jang, Byoung-Gui;Ko, Moon-Suck;Jung, Ha-Yeon;Lee, Sung-Soo
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.633-640
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    • 2008
  • With a double mismatch primer set designed for amplifying the modified DNA sequence fragments, bovine melanocortin-1 receptor(MC1R) gene encoded in Extension locus which plays a critical role in coat color development was analyzed using polymerase chain reaction mediated restriction fragment length polymorphism(PCR-RFLP). Amplified PCR fragments were successfully discriminated with combining the MspI- and AluI-RFLP into three major alleles(ED, E+, and e), directly related to bovine coat color phenotypes. The genotyping results showed that Jeju black cattle contained three MC1R alleles, but yellowish-red colored Hanwoo and bridle colored Korean Brindle cattle did not contained the dominant black allele ED. However, two dominant black-colored cattle breeds, Holstein and Angus, contained the ED allele over 96% in frequency. Hanwoo×Holstein F1 and Hanwoo×Angus F1 crossbred calves showed ED/e MC1R genotypes, and uniformly black coat color. the results suggested that this MC1R genotyping method be useful in allele discrimination for bovine MC1R gene which used for breed classification and characterization, as one of the important genetic markers, using combination of MspI- and AluI-RFLP for modified PCR product amplified with a newly designed double mismatch primer set.

Helicobacter pylori Strain 51 (Korean Isolate): Ordered Overlapping BAC Library, Combined Physical and Genetic Map, and Comparative Analysis with H. pylori Strain 26695 and Strain J99

  • KANG HYUNG-LYUN;LEE WOO-KON;SONG JAE-YOUNG;CHOI SANG-HAENG;PARK SEONG-GYU;RYU BOK-DEOK;LEE EUN-JOO;KIM JI-SUN;PARK JEONG-UCK;BAIK SEUNG-CHUL;CHOI MYOUNG-BUM;YOUN HEE-SHANG;KO GYUNG-HYUCK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.844-854
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    • 2005
  • We constructed a defined physical and genetic map of H. pylori strain 51, previously isolated from a Korean patient with a duodenal ulcer, by combining a restriction analysis by pulse-field gel electrophoresis with the construction of a BAC library. A Notl-digest of H. pylori strain 51 genome yielded seven fragments, from which the genomic size was estimated to be 1,698$\pm$24 kb. The BAC library was constructed from 50 to 200 kb fragments of HindIII-digested genomic DNA. From 700 BAC clones, an ordered overlapping maxi-set of 82 BAC clones was assembled that covered the entire genome. The positions of 15 genes were localized in the strain 51 genome with 4-22 kb of resolution and were compared with their orthologues in strain 26695 and strain J99. The arrangement of the 15 genes was identical in strain 51 and strain J99, except for flaA and hpaA. The plasticity zone of strain 51, like that of strain J99, was located in the single region, and was shorter than those of strain 26695 and strain J99. The strain 51 plasticity zone consisted of ORFs common only to strain 51 and J99 or to strain 51 and 26695, as well as strain 51-specific ORFs. Three genetic translocations and/or inversions were found between orthologue ORFs in strain 51 and strain J99. These results show that the chromosomal organization of strain 51 differs from Western strains such as strain 26695 and strain J99.

Single Nucleotide Polymorphism Exploring the 5'-Regions of Estrogen Receptor-${\alpha}$ Gene and Association With Reproduction Performance and Milk Yield in Hanwoo and Holstein Dairy Cattle (Estrogen Receptor-${\alpha}$ 유전자 5' 영역의 Single Nucleotide Polymorphism의 탐색과 한우와 Holstein에서 번식능력 및 산유능력과의 관계)

  • Yeom, Gyu-Tae;Jeon, Hyang-A;Park, Hae-Geum;Kim, Young Sin;Kim, Hyun;Kim, Jae Hwan;Seong, Hwan-Hoo;Cho, Young Moo;Cho, Jae-Hyeon;Ko, Yeoung-Gyu
    • Reproductive and Developmental Biology
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    • v.38 no.3
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    • pp.123-127
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    • 2014
  • This study was conducted for SNPs in the 5'-regions of estrogen receptor-${\alpha}$ (ESR-${\alpha}$), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genetic improvement was incurred low reproduction performance. The objective of this study was to investigate connections between single nucleotide polymorphisms (SNP) of Estrogen receptor-${\alpha}$ (ESR-${\alpha}$) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to genotype the cattle. The result showed two variants of the ESR-${\alpha}$ gene. The Bgl I cut the 492 bp amplification product into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction fragments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in these breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was $382.18{\pm}10.03$ days, and G/G was $381.69{\pm}5.22$ days. The SPC of A/G was $1.62{\pm}0.16$, and G/G was $1.32{\pm}0.04$. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was $7,253.00{\pm}936.00kg$ and of G/G was $8,747.51{\pm}204.88kg$, showing no significant difference.

Polymorphisms in Heat Shock Proteins A1B and A1L (HOM) as Risk Factors for Oesophageal Carcinoma in Northeast India

  • Saikia, Snigdha;Barooah, Prajjalendra;Bhattacharyya, Mallika;Deka, Manab;Goswami, Bhabadev;Sarma, Manash P;Medhi, Subhash
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8227-8233
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    • 2016
  • Background: To investigate polymorphisms in heat shock proteins A1B and A1L (HOM) and associated risk of oesophageal carcinoma in Northeast India. Materials and Methods: The study includes oesophageal cancer (ECA) patients attending general outpatient department (OPD) and endoscopic unit of Gauhati Medical College. Patients were diagnosed based on endoscopic and histopathological findings. Genomic DNA was typed for HSPA1B1267 and HSPA1L2437 SNPs using the polymerase chain reaction with restriction fragment length polymorphisms. Results: A total of 78 cases and 100 age-sex matched healthy controls were included in the study with a male: female ratio of 5:3 and a mean age of $61.4{\pm}8.5years$. Clinico-pathological evaluation showed 84% had squamous cell carcinoma and 16% were adenocarcinoma. Dysphagia grades 4 (43.5%) and 5 (37.1%) were observed by endoscopic and hispathological evaluation. The frequency of genomic variation of A1B from wild type A/A to heterozygous A/G and mutant G/G showed a positive association [chi sq=19.9, p=<0.05] and the allelic frequency also showed a significant correlation [chi sq=10.3, with cases vs. controls, OR=0.32, $p{\leq}0.05$]. The genomic variation of A1L from wild T/T to heterozygous T/C and mutant C/C were found positively associated [chi sq=7.02, p<0.05] with development of ECA. While analyzing the allelic frequency, there was no significant association [chi sq=3.19, OR=0.49, p=0.07]. Among all the risk factors, betel quid [OR=9.79, Chi square=35.0, p<0.05], tobacco [OR=2.95, chi square=10.6, p<0.05], smoking [OR=3.23, chi square=10.1, p<0.05] demonstrated significant differences between consumers vs. non consumers regarding EC development. Alcohol did not show any significant association [OR=1.34, chi square=0.69, p=0.4] independently. Conclusions: It can be concluded that the present study provides marked evidence that polymorphisms of HSP70 A1B and HSP70 A1L genes are associated with the development of ECA in a population in Northeast India, A1B having a stronger influence. Betel quid consumption was found to be a highly significant risk factor, followed by smoking and tobacco chewing. Although alcohol was not a potent risk factor independently, alcohol consumption along with tobacco, smoking and betel nut was found to contribute to development of ECA.

Association of Leptin Receptor Lys109Arg and Gln223Arg Polymorphisms with Increased Risk of Clear Cell Renal Cell Carcinoma

  • Mu, Hui-Jun;Zou, Jian;Xie, Ping;Xu, Zhuo-Qun;Ruan, Jun;Yang, Shu-Dong;Yin, Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.10
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    • pp.4211-4215
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    • 2014
  • Background: Although roles of genetic polymorphisms of leptin receptor (LEPR) gene in several cancers have been documented, the association between polymorphisms of LEPR and clear cell renal cell carcinoma (CC-RCC) remains unknown. The aim of this study was to explore any relation. Materials and Methods: The study population consisted of 77 patients with CC-RCC and 161 healthy control subjects. Polymorphism analyses of Lys109Arg and Gln223Arg were performed by direct DNA sequencing and PCR-restriction fragment length polymorphism approaches respectively. Results: Comparisons of allelic and genotypic frequencies in Lys109Arg and Gln223Arg showed no significant difference between the cases and controls. However, when evaluating the combined genotype of Lys109Arg and Gln223Arg, risk with GG/GG was increased (OR=1.85, 95%CI=1.04-3.30) and with GA/GG or GG/GA was decreased (OR=0.07, 95%CI=0.01-0.54; OR and 95%CI of the latter could not be calculated for a value of zero). Furthermore, the G-G haplotype frequency of Lys109Arg and Gln223Arg in the cases was higher (OR=1.68; 95%CI=1.02-2.76). In contrast, the A-G and G-A haplotype frequencies in the cases were lower than those in the controls (OR=0.06; 95%CI=0.01 to 0.47; OR and 95%CI of the latter could not be calculated for a value of zero). In addition, the Lys109Arg A allele was in LD with the Gln223Arg A allele (d'=0.9399) in the CC-RCC subjects, but not in the controls. Conclusions: Our data suggest that the GG/GG combined genotype and G-G haplotype of Lys109Arg and Gln223Arg can act as evaluating factors for CC-RCC risk.

Association of Insulin Receptor Substrate-1 G972R Variant with Non-small Cell Lung Cancer Risk

  • Lee, Chang Youl;Ahn, Chul Min;Jeon, Jeong Hee;Kim, Hyung Jung;Kim, Se Kyu;Chang, Joon;Kim, Sung Kyu;Chang, Yoon Soo
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.1
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    • pp.8-13
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    • 2009
  • Background: The insulin receptor substrate-1 (IRS-1) is the primary docking molecule for the insulin-like growth factor I receptor (IGF-IR), and is required for activation of the phosphatidylinositol 3'-kinase (PI3K) pathway. IRS-1 activation of the (PI3K) pathway regulates IGF-mediated survival, enhancement of cellular motility and apoptosis. Therefore, we attempted to ascertain whether IRS-1 genetic variations affect an individual's risk for non-small cell lung cancer (NSCLC). Methods: Two-hundred and eighteen subjects, either diagnosed with NSCLC or control subjects, were matched by age, gender and smoking status. Genomic DNA from each subject was amplified by PCR and analyzed according to the restriction fragment length polymorphism (RFLP) profile to detect the IRS-1 G972R polymorphism. Results: The frequencies of each polymorphic variation, in the control population, were as follows: GG=103 (94.5%) and GR=6 (5.5%); for the NSCLC subjects, the genotypic frequencies were as follows: GG=106 (97.2%) and GR=3 (2.8%). We could not demonstrate statistically significant differences in the genotypic distribution between the NSCLC and the control subjects (p=0.499, Fisher's Exact test). The relative risk of NSCLC, associated with the IRS-1 G972R polymorphic variation, was 1.028 (95% CI; 0.63~9.90). In addition, we found no differences between polymorphic variants with regard to the histological subtype of NSCLC. Conclusion: We did not observe any noteworthy differences in the frequency of the IRS-1 G972R polymorphism in NSCLC patients, compared to control subjects. These results suggest suggesting that, in our study population, the IRS-1 G972R polymorphism does may not appear to be associated with an increased risk of NSCLC.

An Association of C/T Polymorphism in the TFF1 Gene and the Susceptibility to Gastric Cancer (TFF1 유전자의 C/T 다형성과 위암 민감성과의 연관성)

  • Maeng, Eun Jae;Song, Jae Hwi;Sung, Soo Yoon;Cao, Zhang;Park, Won Sang
    • Journal of Gastric Cancer
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    • v.8 no.3
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    • pp.113-119
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    • 2008
  • Purpose: This study investigated whether a single nucleotide polymorphism (SNP) located at position -2 in the Kozak sequence of the TFF1 gene is associated with H. pylori infection and the development of gastric cancer in Koreans. Materials and Methods: We enrolled 167 patients with gastric cancer from January 2000 to December 2003 and also 299 healthy controls during the same period. The genotype of the TFF1 SNP was analyzed by polymerase chain reaction-restriction fragment length polymorphism and single strand conformation polymorphism. We also examined the H. pylori infection by Giemsa staining. Results: No significant difference in the allele or the TFF1 SNP genotype frequency was observed between the patients with gastric cancer and the control subjects (P=0.595 and P=0.715, respectively). When stratified by the histological subtype of gastric cancer and the age of the patients, the risk was not statistically significant between the two study groups (P=0.088 and P=0.551, respectively). H. pylori infection was detected in 39 cases and it was not associated with the TFF1 genotype. Conclusion: These findings suggest that this TFF1 gene polymorphism is not associated with H. pylori infection and gastric cancer in Koreans and so it doesn't contribute to the susceptibility to gastric cancer in Koreans.

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