• Title/Summary/Keyword: DNA variants

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Endoreduplication Pattern of Somatic Embryos and Variants Occurrence Affected by Pre-existed Endoreduplicated Cells in Doritaenopsis (Doritaenopsis 체세포배의 내배수성 특성과 절편체의 내배수성 세포에 기인한 체세포변이의 발생)

  • Park, So-Young;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.297-302
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    • 2006
  • In general, the proliferation of orchids via somatic embryos has been used for mass production of somatic clones because of high propagation efficiency. In spite of high propagation rate, this method often brings somaclonal variation, especially polyploid frequency. Therefor we here concentrated to investigate the relationship between endopolyploidization patterns of explants and the occurrence of tetraploid variant in clonally proliferated Doritaenopsis via somatic embryo regeneration system. In the fully developed somatic embryo, upper part contained 2C to 16C while middle and lower parts showed 2C to 32C DNA content. Two-week-old embryo contained 2C to 16C, whereas those regenerated after 4 to 10-week-old contained 2C to 64C nuclei. Results showed that endoreduplication was variable depending upon tissue types, ages, and parts in one species. lower part of somatic embryo having high endoreduplication degree increased the regeneration of tetraploid variants by about 3-fold comparing to upper part of somatic embryo culture. polyploid frequency occurrence might be closely related to the high levels of endoreduplication of somatic embryos used as explant. It suggested that the upper part of somatic embryo having comparatively low endoreduplication degree is suitable for the stable in vitro propagation system.

Molecular Cloning of Glycoside Hydrolase Family 74 Genes and Analysis of Transcript Products from the Basidiomycete Phanerochaete chrysosporium (담자균 Phanerochaete chrysosporium으로부터 유래한 Glycoside Hydrolase Family 74 유전자 클로닝과 전사산물 분석)

  • Lee, Jae-Won;Samejima, Masahiro;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.3
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    • pp.56-63
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    • 2006
  • In order to evaluate the mechanism of cellulose hydrolysis, the complementary DNA encoding Glycoside Hydrolase Family (GHF)74 was cloned from Phanerochaete chrysosporium. Depending on the presence of Cellulose Binding Module (CBM), it can be classified as GHF74A or GHF74B. The GHF74A gene from P. chrysosporium (PcGHF74A) consists of 2163 bp encoding a protein of 721 amino acid residues. The PcGHF74A showed homology of 70~77% compared with the GHF74 from other filamentous fungi. The PcGHF74B, which contains CBM and is a member of family 1, was transcribed to various transcripts depending on the nature of carbon sources and their concentration. To study the possible presence of splice variants in GHF74B transcripts in P. chrysospoium, we carried out RT-PCR analysis using primers that designed based on the annotation data and sequenced data. Our result indicated that PcGHF74B was transcribed to several splicing variants in various culture conditions. Especially in the culture of 2% cellulose, three transcript products were observed. First transcript was presumed to be a full length ORF that contained 11th intron with stop codon at position 2562 bp. The second one consisted of 12 exons and 11 introns with stop codon at position 1187 bp with 7th exon. The shortest transcript consisted of 10 exons and 9 introns with stop codon at 910 bp in the 7th exon. These premature stop codon might prevent the synthesis of fully active GHF74 or contribute for the production of protein with distinct function depending on the ambient carbon sources.

Prevalence of Transfusion Transmitted Virus-Like Mini Virus in Children (소아의 Transfusion Transmitted Virus-Like Minivirus 유병률)

  • Chung, Ju Young;Han, Tae Hee
    • Pediatric Infection and Vaccine
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    • v.11 no.2
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    • pp.153-157
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    • 2004
  • Purpose : Ttransfusion transmitted virus(TTV) is a circular DNA and consists of diverse genotypes and variants. The pathogenecity of TTV is still unclear. Recently another circular single stranded DNA virus, distantly related to TTV was isolated from the sera of blood donors, designated as Transfusion transmitted virus like minivirus(TLMV). TTV and TLMV show greater sequence divergence from each other than between genotypes of TTV. We planned to know the prevalence of TLMV in children. Methods : TLMV DNA was detected by PCR primers from noncoding region of the genome in 88 children without hepatitis, aged 0~15 years. PCR products derived from 10 children were directly sequenced and phylogenetic analysis was undertaken. Results : TLMV DNA was detected in 49% of 88 children without hepatitis. The prevalence of TLMV varied with age : <1 y, 16%(4/25); 1~3 y, 62%(18/29); 4~6 y, 43%(7/16); 7~9 y. 16%(1/6); 10~15 y, 66%(8/12). Mixed infection with TTV was confirmed in 22% of 88 children. Pyhlogenetic analysis of 10 TLMV sequences showed much heterogeneity compared to sequences of GenBank. Conclusion : TLMV prevalence in children was 49% in Korean children. Our TLMV sequence did not cluster in any sequence of TLMV in the GenBank.

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Screening of genes differentially expressed in cultured human periodontal ligament cells and human gingival fibroblasts (배양된 치주인대세포와 치은섬유아세포에서 상이하게 발현된 유전자들의 검토 양상)

  • Yoon, Hye-Jeong;Choi, Mi-Hye;Yeo, Shin-II;Park, Jin-Woo;Choi, Byung-Ju;Kim, Moon-Kyu;Kim, Jung-Chul;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.36 no.3
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    • pp.613-625
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    • 2006
  • Periodontal ligament(PDL) cells and human gingival fibroblasts(HGFs) play important roles in development, regeneration, normal function, and pathologic alteration. PDL cells and HGFs have the similarity related with general characteristics of fibroblast such as spindle shaped morphology, the presence of vimentin intermediate filament and the synthesis of interstitial collagens and fibronectin. There were many studies about the differences between PDL cells and HGFs, but they were not about whole gene level. In this study, we tried to explain the differences of gene expression profiles between PDL cells and HGFs, and the differences among three individuals by screening gene expression patterns of PDL cells and HGFs, using cDNA microarray. Although there were some variants among three experiments, a set of genes were consistentely and differentially expressed in one cell type. Among 3,063 genes, 49 genes were more highly expressed in PDL cells and 12 genes were more highly expressed in HGFs. The genes related with cell structure and motility were expressed more highly in PDL cells. These are cofilin 1, proteoglycan 1 secretory granule, collagen type I(${\alpha}$ 1), adducin gamma subunit, collagen type III(${\alpha}$ 1), fibronectin, lumican(keratan sulfate proteoglycan), and ${\alpha}$ -smooth muscle actin. Tissue inhibitor of metalloproteinase known as the enzyme controlling extracellular matrix with matrix metalloproteinase is more highly expressed in PDL cells, osteoprotegerin known as osteoclastogenesis inhibitory factor is more highly expressed in HGFs. We performed northern blot to verify cDNA microarray results on selected genes such as tissue inhibitor of metalloproteinase, fibronectin, osteoprogeterin. The result of northern blot analysis showed that each cell expressed the genes in similar pattern with cDNA microarray result. This result indicates that cDNA microarray is a reliable method in screening of gene expression profiles.

Association of a miR-502-Binding Site Single Nucleotide Polymorphism in the 3'-Untranslated Region of SET8 and the TP53 Codon 72 Polymorphism with Cervical Cancer in the Chinese Population

  • Yang, Shao-Di;Cai, Yan-Lin;Jiang, Pei;Li, Wen;Tang, Jian-Xin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6505-6510
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    • 2014
  • Objective: This study was conducted to identify whether polymorphic variants of set domain-containing protein 8 (SET8) and tumor protein p53 (TP53) codon 72, either independently or jointly, might be associated with increased risk for cervical cancer. Methods: We genotyped SET8 and TP53 codon 72 polymorphisms of peripheral blood DNA from 114 cervical cancer patients and 200 controls using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct DNA sequencing. Results: The frequency of SET8 CC (odds ratios (OR) = 2.717, 95% CI=1.436-5.141) or TP53 GG (OR=2.168, 95% CI=1.149-4.089) genotype was associated with an increased risk of cervical cancer on comparison with the SET8 TT or TP53 CC genotypes, respectively. In additional, interaction between the SET8 and TP53 polymorphisms increased the risk of cervical cancer in a synergistic manner, with the OR being 9.913 (95% CI=2.028-48.459) for subjects carrying both SET8 CC and TP53 GG genotypes. Conclusion: These data suggest that there are significant associations between the miR-502-binding site SNP in the 3'-UTR of SET8 and the TP53 codon 72 polymorphism with cervical cancer in Chinese, and there is a gene-gene interaction.

RB1 Polymorphism Contributes to the Efficacy of Platinum-Taxanes in Advanced Squamous Cell Lung Cancer

  • Liu, Di;Xu, Wen;Zhang, Zhi-Wei;Qian, Ji;Zheng, Hui;Zhang, Jie;Su, Bo
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.775-781
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    • 2015
  • Background: RB1 (retinoblastoma 1) was reportedly one of the major determinative factors for sensitivity to taxanes in previous studies. In this study, we investigated the influence of RB1 single nucleotide polymorphisms (SNPs) on the efficacy of platinum-taxane regimens in advanced NSCLC patients. Materials and Methods: 234 cases of patients with advanced NSCLC who were treated with first-line platinum-taxane agents were enrolled in this study. Genomic DNA was extracted from patients' peripheral blood samples using a QIAamp DNA Maxi Kit, and genotyped by iSelect HD Bead-Chip. Results: Regression analyses were conducted through the univariate and multivariate Cox proportional hazards model in the 234 patients. The results showed that of the eight RB1 tagSNPs, only rs4151510 was a positive predictive factor for the advanced NSCLC patients treated with platinum taxanes regimen. The patients with G/G genotype of RB rs4151510 had longer overall survival (OS) than the non-G/G genotype (p=0.018). The histology was also correlated with OS in the whole advanced NSCLC patients. Three tagSNPs of RB1, rs4151510, rs4151465, rs9568036 were significantly associated with OS in the advanced NSCLC patients with squamous cell histology using Kaplan-Meier overall survival analysis stratified by histology. Conclusions: RB1 genomic variants were correlated with the efficacy of platinum-taxanes regimen. RB rs4151510 is an independent factor of the prognosis of NSCLC patients receiving platinum-taxane chemotherapy.

Genetic Screening for Mutations in the Chip Gene in Intracranial Aneurysm Patients of Chinese Han Nationality

  • Su, Li;Zhang, Yuan;Zhang, Chun-Yang;Zhang, An-Long;Mei, Xiao-Long;Zhao, Zhi-Jun;Han, Jian-Guo;Zhao, Li-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1687-1689
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    • 2013
  • We performed a case-control study to investigate whether SNPs of CHIP might affect the development of IA in Chinese Han nationality. We believe we are the first to have screened IA patients for mutations in the CHIP gene to determine the association with these variants. The study group comprised 224 Chinese Han nationality patients with at least one intracranial aneurysm and 238 unrelated healthy Han nationality controls. Genomic DNA was isolated from blood leukocytes. The entire coding regions of CHIP were genotyped by PCR amplification and DNA sequencing. Differences in genotype and allele frequencies between patients and controls were tested by the chi-square method. Genotype and allele frequencies of the SNP rs116166850 was demonstrated to be in Hardy-Weinberg equilibrium. No significant difference in genotype or allele frequencies between case and control groups was detected at the SNP. Our data do not support the hypothesis of a major role for the CHIP gene in IA development in the Chinese Han population.

Evaluation of Risk Factors for Nasopharyngeal Carcinoma in a High-risk Area of India, the Northeastern Region

  • Lourembam, Deepak Singh;Singh, Asem Robinson;Sharma, T. Dhaneshor;Singh, Th Sudheeranjan;Singh, Thiyam Ramsing;Singh, Lisam Shanjukumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.12
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    • pp.4927-4935
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    • 2015
  • Northeastern India is a major nasopharyngeal carcinoma (NPC) high risk-area although the rest of the country has very low incidence. A case-control study of 105 NPC cases and 115 controls was conducted to identify the potential risk factors for NPC development in this region. Information was collected by interviewer about socio-demographic characteristics, cigarette smoking, alcohol consumption, dietary history, occupational history, and a family history of cancer. Epstein-Barr viral load was assayed from the blood DNA by real time PCR. Associations between GSTs genotypes, cytochrome P450 family including CYP1A1, CYP2E1 and CYP2A6 polymorphisms and susceptibility to relationship between the diseases were studied using PCR-RFLP assay. Results indicate that Epstein-Barr virus load was significantly higher in patients compared to controls (p<0.0001). Furthermore, concentration of blood EBV-DNA was significantly higher in advanced stage disease (Stage III and IV) than in early stage disease (Stage I and II) (p<0.05). Presence of CYP2A6 variants that reduced the enzyme activity was significantly less frequent in cases than controls. Smoked meat consumption, exposure to smoke, living in poorly ventilated house and alcohol consumption were associated with NPC development among the population of Northeastern India. Thus, overall our study revealed that EBV viral load and genetic polymorphism of CYP2A6 along with living practices which include smoked meat consumption, exposure to smoke, living in poorly ventilated houses and alcohol consumption are the potential risk factors of NPC in north eastern region of India. Understanding of the risk factors and their role in the etiology of NPC are helpful forpreventive measures and screening.

Role of DNA Repair-related Gene Polymorphisms in Susceptibility to Risk of Prostate Cancer

  • Yang, Bo;Chen, Wei-Hua;Wen, Xiao-Fei;Liu, Hui;Liu, Feng
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5839-5842
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    • 2013
  • Aim: We assessed the association between genetic variants of XPG, XPA, XPD, CSB, XPC and CCNH in the nucleotide excision repair (NER) pathway and risk of prostate cancer. Methods: We genotyped the XPG, XPA, XPD, CSB, XPC and CCNH polymorphisms by a 384-well plate format on the MassARRAY(R) platform. Multivariate logistical regression analysis was used to assess the associations between the six gene polymorphisms and risk of prostate cancer. Results: Individuals carrying the XPG rs229614 TT (OR=2.01, 95%CI=1.35-3.27) genotype and T allele (OR=1.73, 95%CI=1.37-2.57) were moderately significantly associated with a higher risk of prostate cancer. Subjects with XPD rs13181 G allele had a marginally increased risk of prostate cancer, with adjusted OR(95%CI) of 1.53 (1.04-2.37). Moreover, individuals carrying with CSB rs2228526 GG genotype (OR=2.05, 95% CI=1.23-3.52) and G allele (OR=1.56, 95%CI=1.17-2.05) were associated with a higher increased risk of prostate cancer. The combination genotype of XPG rs2296147 T and CSB rs2228526 G allele had accumulative effect on the risk of this cancer, with an OR (95% CI) of 2.23(1.37-3.59). Conclusions: Our study indicates that XPG rs2296147 and CSB rs2228526 polymorphisms are significantly associated with increased risk of prostate cancer, and that combination of XPG rs2296147 T allele and CSB rs2228526 G allele is strongly associated with an increased risk.

Cryopreservation of Embryogenic Tissue and Plant Regeneration in Larix leptolepis (낙엽송 (Larix leptolepis) 배발생조직의 초저온보존 및 식물체 재분화)

  • Kim, Yong-Wook;Kim, Joon-Chul;Youn, Yang;Noh, Eu-Rae;Son, Sung-Ho
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.229-233
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    • 1999
  • The possibility for long-term preservation of Larix leptolepis embryogenic tissue was tested in this study. Higher relative increase of the tissue fresh weight was observed when embryogenic tissue was pretreated for 24 hrs in a medium containing 0.4 M sorbitol or 20% polyethyleneglycol with cooling rate of -0.33$^{\circ}C$/min. The fast cooling rate of -0.5$^{\circ}C$ and -1.$0^{\circ}C$/min appeared to be less effective in regrowth of tissues from cryopreservation. No DNA variants have been observed by PCR analysis among the embryogenic tissues recovered after 1-, 7-, and 28-day-cryopreservation. The post-thaw embryogenic tissue gave rise to mature somatic embryos which developed into plants.

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