• Title/Summary/Keyword: Materials genome

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Situation of HPV16 E2 Gene Status During Radiotherapy Treatment of Cervical Carcinoma

  • Kahla, Saloua;Kochbati, Lotfi;Maalej, Mongi;Oueslati, Ridha
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2869-2873
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    • 2014
  • Background: Human papillomavirus (HPV) integration within the E2 gene has been proposed as a critical event in cervical carcinogenesis. This study concerned whether HPV16 status and E2 gene intactness are predictive of radiation response in patients with cervical cancer. Materials and Methods: Biopsies of 44 patients with cervical cancer were collected before or after radiotherapy. The presence of HPV16 was assessed by polymerase chain reaction (PCR) using specific primers for the L1 region. E2 disruption was detected by amplifying the entire E2 gene. Results: HPV16 DNA was found in 54.5% of the clinical samples. Overall, 62.5% of the HPV16 positive tumors had integrated viral genome and 37.5% had episomal genome. There was a tendency of increase of HPV16 E2 negative tumors compared with HPV16 L1 ones in advanced stages (75% versus 20% in stage III respectively). Detection of E2 gene appeared influenced by the radiotherapy treatment, as the percentage of samples containing an intact HPV16 E2 was more frequent in pretreated patients compared to radiotherapy treated patients (66.6% versus 20%). The radiation therapy caused an eight-fold [OR= 8; CI=1.22-52.25; p=0.03] increase in the risk of HPV16 genome disruption. The integration status is influenced by the irradiation modalities, interestingly E2 disruption being found widely after radiotherapy treatment (75%) with a total fractioned dose of 50Gy. Conclusions: This study reveals that the status of the viral DNA may be used as a marker to optimize the radiation treatment.

Identification of Human LRG1 Polymorphisms and Their Genetic Association with Rheumatoid Arthritis

  • Jin, Eun-Heui;Chae, Soo-Cheon;Shim, Seung-Cheol;Kim, Hwan-Gyu;Chung, Hun-Taeg
    • Genomics & Informatics
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    • v.6 no.2
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    • pp.77-83
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    • 2008
  • Human leucine-rich alpha-2-glycoprotein 1 (LRG1) was first identified as a trace protein in human serum. The primary sequence of LRG1 includes repeated leucine residues and putative membrane-binding domains. But, there is no published information on the genetic variation of this gene. In this study, LRG1 was identified as one of several upregulated genes in RA patients. We examined the expression levels of LRG1 between an RA patient and a healthy control by RT-PCR and validated that LRG1 was highly expressed in RA patients compared with controls. We identified the possible variation sites and single nucleotide polymorphisms (SNPs) in the human LRG1 gene by direct sequencing and analyzed the association of genotype and allele frequencies between RA patients and a control group without RA. We further investigated the relationship between these polymorphisms and the level of RF or anti-CCP in RA patients. We identified a total of three SNPs(g.-678A>G, g.-404C>T and g.1427T>C) and two variation sites (g.-1198delA and g.-893delA) in the LRG1 gene. Our results suggest that polymorphisms of the LRG1 gene are not associated with the susceptibility of RA in the Korean population.

Microbial Biotechnology Powered by Genomics, Proteomics, Metabolomics and Bioinformatics

  • Lee, Sang-Yup
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2000.11a
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    • pp.13-16
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    • 2000
  • Microorganisms have been widely employed for the production of useful bioproducts including primary metabolites such as ethanol, succinic acid, acetone and butanol, secondary metabolites represented by antibiotics, proteins, polysaccharides, lipids and many others. Since these products can be obtained in small quantities under natural condition, mutation and selection processes have been employed for the improvement of strains. Recently, metabolic engineering strategies have been employed for more efficient production of these bioproducts. Metabolic engineering can be defined as purposeful modification of cellular metabolic pathways by introducing new pathways, deleting or modifying the existing pathways for the enhanced production of a desired product or modified/new product, degradation of xenobiotics, and utilization of inexpensive raw materials. Metabolic flux analysis and metabolic control analysis along with recombinant DNA techniques are three important components in designing optimized metabolic pathways, This powerful technology is being further improved by the genomics, proteomics, metabolomics and bioinformatics. Complete genome sequences are providing us with the possibility of addressing complex biological questions including metabolic control, regulation and flux. In silico analysis of microbial metabolic pathways is possible from the completed genome sequences. Transcriptome analysis by employing ONA chip allows us to examine the global pattern of gene expression at mRNA level. Two dimensional gel electrophoresis of cellular proteins can be used to examine the global proteome content, which provides us with the information on gene expression at protein level. Bioinformatics can help us to understand the results obtained with these new techniques, and further provides us with a wide range of information contained in the genome sequences. The strategies taken in our lab for the production of pharmaceutical proteins, polyhydroxyalkanoate (a family of completely biodegradable polymer), succinic acid and me chemicals by employing metabolic engineering powered by genomics, proteomics, metabolomics and bioinformatics will be presented.

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Alu Hypomethylation in Smoke-Exposed Epithelia and Oral Squamous Carcinoma

  • Puttipanyalears, Charoenchai;Subbalekha, Keskanya;Mutirangura, Apiwat;Kitkumthorn, Nakarin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5495-5501
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    • 2013
  • Background: Alu elements are one of the most common repetitive sequences that now constitute more than 10% of the human genome and potential targets for epigenetic alterations. Correspondingly, methylation of these elements can result in a genome-wide event that may have an impact in cancer. However, studies investigating the genome-wide status of Alu methylation in cancer remain limited. Objectives: Oral squamous cell carcinoma (OSCC) presents with high incidence in South-East Asia and thus the aim of this study was to evaluate the Alu methylation status in OSCCs and explore with the possibility of using this information for diagnostic screening. We evaluated Alu methylation status in a) normal oral mucosa compared to OSCC; b) peripheral blood mononuclear cells (PBMCs) of normal controls comparing to oral cancer patients; c) among oral epithelium of normal controls, smokers and oral cancer patients. Materials and Methods: Alu methylation was detected by combined bisulfite restriction analysis (COBRA) at 2 CpG sites. The amplified products were classified into three patterns; hypermethylation ($^mC^mC$), partial methylation ($^uC^mC+^mC^uC$), and hypomethylation ($^uC^uC$). Results: The results demonstrate that the $%^mC^mC$ value is suitable for differentiating normal and cancer in oral tissues (p=0.0002), but is not significantly observe in PBMCs. In addition, a stepwise decrease in this value was observed in the oral epithelium from normal, light smoker, heavy smoker, low stage and high stage OSCC (p=0.0003). Furthermore, receiver operating characteristic (ROC) curve analyses demonstrated the potential of combined $%^mC$ or $%^mC^mC$ values as markers for oral cancer detection with sensitivity and specificity of 86.7% and 56.7%, respectively. Conclusions: Alu hypomethylation is likely to be associated with multistep oral carcinogenesis, and might be developed as a screening tool for oral cancer detection.

Monitoring of Red Pepper Powder and Seasoned Red-Pepper Sauce using Species-Specific PCR in Conjunction with Whole Genome Amplification

  • Hong, Yewon;Kwon, Kisung;Kang, Tae Sun
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.146-150
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    • 2018
  • Red pepper is one of the most important spices popularly utilized in Korea. Because of the differences in tariff rates between red pepper powder and seasoned red-pepper sauce, seasoned red-pepper sauce is often therefore imported by consumers, then dried, ground, and added to red pepper powder for cost effective purposed to use the product the most effectively. In this study, we combined species-specific polymerase chain reaction (PCR) assays (for red pepper, garlic, onion, spring onion, and ginger) with whole-genome amplification (WGA). Thirty-nine red pepper powders were well in accordance with their labels. However, six red pepper powder and five seasoned red-pepper sauce products failed to meet their compliance requirements. As a consequence, our monitoring results revealed that the overall mislabeling rate detected in this study was identified at 22%. Thus, our findings showed that the species-specific PCR in conjunction with WGA was an ideal method to identify raw materials that are used in the manufacturing of red pepper powder and seasoned red-pepper sauce.

High-Throughput Development of Polymorphic Simple Sequence Repeat Markers Using Two Whole Genome Sequence Data in Peucedanum japonicum

  • Lee, Junki;Joh, Ho Jun;Kim, Nam-Hoon;Lee, Sang-Choon;Jang, Woojong;Choi, Beom Soon;Yu, Yeisoo;Yang, Tae-Jin
    • Plant Breeding and Biotechnology
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    • v.5 no.2
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    • pp.134-142
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    • 2017
  • Resource plants are important and have strong potential for a variety of utilities as crops or pharmaceutical materials. However, most resource plants remain wild and thus their utility for breeding and biotechnology is limited. Molecular markers are useful to initiate genetic study and molecular breeding for these understudied resource plants. We collected various wild collections of Peucedanum japonicum which is indigenous resource plants utilized as oriental medicine and leafy vegetables in Korea. In this study, we produced two independent whole genome sequences (WGSs) from two collections and identified large scale polymorphic simple sequence repeat (pSSR) based on our pipeline to develop SSR markers based on comparison of two WGSs. We identified a total of 452 candidate pSSR contigs. To confirm the accuracy and utility of pSSR, we designed ten SSR primer pairs and successfully applied those to seven collections of P. japonicum. The WGS and pSSR candidates identified in this study will be useful resource for genetic research and breeding purpose for the valuable resource plant, P. japonicum.

In silico Analysis of PERVs Based on the Porcine Genomic Sequence Information (돼지 유전체 염기서열을 이용한 내인성 리트로 바이러스 분석에 관한 연구)

  • Yu, Seong-Lan;Lee, Jun-Heon
    • Korean Journal of Agricultural Science
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    • v.36 no.2
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    • pp.159-165
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    • 2009
  • This study was conducted to identify the PERV (Porcine Endogenous Retrovirus) integration sites and their characterizations using the porcine genomic sequence information. Total 114 Mb (4.2%) sequence of the 2.7 Gb pig genome was investigated for the PERV sequences. As the results, 8 PERV sequences were identified and their genomic structures were deduced from the BLAST searches against previously known PERV genes. Seven PERVs have internal deletions in the protein coding region and they will not be functional. The other one also has internal deletions in the gag and env genes, indicating this PERV is also defective. Even though we could not identify the functional PERVs in this study, the results presented here can be used for the fundamental research materials for controlling PERV infections in relation to xenotransplantation using porcine organs and tissues.

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Assessment of Endophytic Fungal Diversity and Beyond

  • Kim, Soonok
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.20-20
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    • 2015
  • Endophytic fungi are microorganisms inhabiting living plant tissues without causing apparent harm to the host. They are drawing increasing attention due to their ability to produce various bioactive compounds as well as their effects on host growth and resistance to biotic and abiotic stresses. As a first step to assess biodiversity of plant associated fungi in Korea and the following evaluation on diverse biological activities, we are collecting endophytic fungi from plant in wild followed by systematic long-term storage in liquid nitrogen. Molecular identification using ITS sequences was also incorporated for pure culture by hyphal tip isolation. As of April 2015, about 1,400 fungal strains had been isolated from about 170 plant taxa. Fungal isolates belonging to Pleosporales, Diaporthales, Glomerellales, Hypocreales, and Xylariales were the most abundant. These collections are being used for several complementary researches, including screening of isolates with novel bioactive compounds or conferring drought stress resistance, phylogenetic and genomic study. Genome sequencing was performed for 3 isolates, one Xylaria sp. strain JS573 producing griseofulvin, an antifungal compound, and two Fusarium spp. strains JS626 and JS1030, which are assumed to be new species found in Korea. More detailed analysis on these genomes will be presented. These collections and genome informations will serve as invaluable resources for identifying novel bioactive materials in addition to expand our knowledge on fungal biodiversity.

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Systematic Development of Tomato BioResources in Japan

  • Ariizumi, Tohru;Aoki, Koh;Ezura, Hiroshi
    • Interdisciplinary Bio Central
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    • v.3 no.1
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    • pp.1.1-1.6
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    • 2011
  • Recently, with the progress of genome sequencing, materials and information for research on tomato (Solanum lycopersicum) have been systematically organized. Tomato genomics tools including mutant collections, genome sequence information, full-length cDNA and metabolomic datasets have become available to the research community. In Japan, the National BioResource Project Tomato (NBRP Tomato) was launched in 2007, with aims to collect, propagate, maintain and distribute tomato bioresources to promote functional genomics studies in tomato. To this end, the dwarf variety Micro-Tom was chosen as a core genetic background, due to its many advantages as a model organism. In this project, a total of 12,000 mutagenized lines, consisting of 6000 EMS-mutagenized and 6000 gamma-ray irradiated M2 seeds, were produced, and the M3 offspring seeds derived from 2236 EMS-mutagenized M2 lines and 2700 gamma-ray irradiated M2 lines have been produced. Micro-Tom mutagenized lines in the M3 generation and monogenic Micro-Tom mutants are provided from NBRP tomato. Moreover, tomato cultivated varieties and its wild relatives, both of these are widely used for experimental study, are available. In addition to these bioresources, NBRP Tomato also provides 13,227 clones of full-length cDNA which represent individual transcripts non-redundantly. In this paper, we report the current status of NBRP Tomato and its future prospects.