• Title/Summary/Keyword: MOLECULAR MARKER

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Epigenetic Field for Cancerization

  • Ushijima, Toshikazu
    • BMB Reports
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    • v.40 no.2
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    • pp.142-150
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    • 2007
  • Epigenetic alterations, represented by aberrant DNA methylation, are deeply involved in human cancers. In gastric cancers, tumor-suppressor genes are inactivated more frequently by promoter methylation than by mutations. We recently showed that H. pylori infection, a potent gastric carcinogenic factor, induces methylation of specific genes in the gastric mucosae. When the methylation levels were analyzed in the gastric mucosae of healthy volunteers, cases with a single gastric cancer, and cases with multiple gastric cancers, who have increasing levels of risks for gastric cancers, there was a significant increasing trend in the methylation levels among the individuals without current H. pylori infection. This finding unequivocally showed the presence of an epigenetic field for cancerization. The degree of the field defect was measured more conveniently using methylation levels of marker genes than using those of tumor-suppressor genes. The presence of an epigenetic field for cancerization has been indicated for liver, colon, Barrett's esophageal, lung, breast, and renal cancers. Since decreased transcription is involved in the specificity of methylated genes, it is likely that specific genes are methylated according to carcinogenic factors. These findings emphasize the usefulness of DNA methylation as a marker for past exposure to carcinogens and future risk of cancer development.

Conditional Replication of a Recombinant Adenovirus Studied Using Neomycin as a Selective Marker

  • Xue, Feng;Qi, Yi-Peng;Joshua, Mallam Nock;Lan, Ping;Dong, Chang-Yuan
    • BMB Reports
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    • v.36 no.3
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    • pp.275-281
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    • 2003
  • An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated, under identical conditions, that the U251 cells were more sensitive to r2/Ad replication than the EJ cells. In this paper, we report that r2/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus and has great potential in cancer gene therapy.

Intraspecific variation of gene structure in the mitochondrial large subunit ribosomal RNA and cytochrome c oxidase subunit 1 of Pyropia yezoensis (Bangiales, Rhodophyta)

  • Hwang, Il Ki;Kim, Seung-Oh;Hwang, Mi Sook;Park, Eun-Jeong;Ha, Dong-Soo;Lee, Sang-Rae
    • ALGAE
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    • v.33 no.1
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    • pp.49-54
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    • 2018
  • Red algal mitochondrial genomes (mtDNAs) can provide useful information on species identification. mtDNAs of Pyropia / Porphyra (Bangiales, Rhodophyta) have shown diverse variation in their size and gene structure. In particular, the introns and intronic open reading frames found in the ribosomal RNA large subunit gene (rnl) and cytochrome c oxidase subunit 1 gene (cox1) significantly vary the mitochondrial genome size in Pyropia / Porphyra species. In this study, we examined the exon / intron structure of rnl and cox1 genes of Pyropia yezoensis at the intraspecific level. The combined data of rnl and cox1 genes exhibited 12 genotypes for 40 P. yezoensis strains, based on the existence of introns. These genotypes were more effective to identify P. yezoensis strains in comparison to the traditional DNA barcode cox1 marker (5 haplotypes). Therefore, the variation in gene structure of rnl and cox1 can be a novel molecular marker to discriminate the strains of Pyropia species.

High sensitivity of embryonic stem cells to proteasome inhibitors correlates with low expression of heat shock protein and decrease of pluripotent cell marker expression

  • Park, Jeong-A;Kim, Young-Eun;Ha, Yang-Hwa;Kwon, Hyung-Joo;Lee, Young-Hee
    • BMB Reports
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    • v.45 no.5
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    • pp.299-304
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    • 2012
  • The ubiquitin-proteasome system is a major proteolytic system for nonlysosomal degradation of cellular proteins. Here, we investigated the response of mouse embryonic stem (ES) cells under proteotoxic stress. Proteasome inhibitors induced expression of heat shock protein 70 (HSP70) in a concentration- and time-dependent manner, and also induced apoptosis of ES cells. Importantly, more apoptotic cells were observed in ES cells compared with other somatic cells. To understand this phenomenon, we further investigated the expression of HSP70 and pluripotent cell markers. HSP70 expression was more significantly increased in somatic cells than in ES cells, and expression levels of pluripotent cell markers such as Oct4 and Nanog were decreased in ES cells. These results suggest that higher sensitivity of ES cells to proteotoxic stress may be related with lower capacity of HSP70 expression and decreased pluripotent cell marker expression, which is essential for the survival of ES cells.

Genetic Variability Based on Randomly Amplified Polymorphic DNA in Mistletoe Fig (Ficus deltoidea Jack) Collected from Peninsular Malaysia

  • Bhore, Subhash Janardhan;Arneida H., Nurul;Shah, Farida Habib
    • Journal of Forest and Environmental Science
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    • v.25 no.1
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    • pp.57-65
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    • 2009
  • Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

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Development of RAPD-Derived SCAR Markers and Multiplex-PCR for Authentication of the Schisandrae Fructus (오미자 (五味子) 종 감별을 위한 RAPD 유래 SCAR Marker 및 Multiplex-PCR 기법 개발)

  • Lee, Young Mi;Moon, Byeong Cheol;Ji, Yunui;Seo, Hyeong Seok;Kim, Ho Kyoung
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.3
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    • pp.165-173
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    • 2013
  • The fruits of Schisandra chinensis have been used as an edible ingredient and traditional medicine in Korea. Due to morphological similarities of dried mature fruits, the correct identification of S. chinensis from other closely related Schisandrae species is very difficult. Therefore, molecular biological tools based on genetic analysis are required to identify authentic Schisandrae Fructus. Random amplifed polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop an easy, reliable and reproducible method for the authentication of these four species. In this paper, we developed several RAPD-derived species specific SCAR markers and established a multiplex-PCR condition suitable to discriminate each species. These genetic markers will be useful to distinguish and authenticate Schisandrae Fructus and four medicinal plants, S. chinensis, S. sphenanthera, S. repanda and K. japonica, in species level.

Expression and characterization of the recombinant human galectin-3 (유전자 재조합 Human galectin-3의 발현과 성상)

  • Kim, Byung-gyu;Woo, Hee-jong
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.547-554
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    • 1997
  • Galectin-3 is known as an animal ${\beta}$-galactoside-binding lectin charicterized with S-type carbohydrate recognition domain. It plays a role in growth, adherence and movement of cells. It is, also, related to the cell transformation and metastasis of tumor cells. In this study, we have expressed and purified recombinant human galectin-3 (rHgalectin-3) using E coli system and asialofetuin affinity chromatography for the future development of monoclonal antibody to Hgalectin-3, which is suggested as the tumor marker for the gastric and thyroid gland cancers. Expressed protein was confirmed as the Hgalectin-3 by immunoblot with cross-reactive murine monoclonal antibody. Lectin activity and specificity of purified protein were, also, confirmed by the competitive inhibition with galectin-3 specific carbohydrate, lactose. Like physiological galectin-3, lectin activity of the molecule was not changed in nonreduced condition. Dimer formation, furthermore, was observed at high concentration of the protein even in the reduced condition, which is well known in physiological galectin-3. These results showed purified rHgalectin-3 has the same activity and molecular nature compared to the physiological galectin-3.

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Genetic Mapping of Hypernodulation in Soybean Mutant SS2-2

  • Lee, Suk-Ha;Ha, Bo-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.5
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    • pp.416-419
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    • 2001
  • Hypernodulation soybean mutant, SS2-2, is characterized with greater nodulation and nitrogen fixing ability in the root nodule than its wild type, Shinpaldalkong 2. The present study was performed to identify a genetic locus conferring hypernodulation in soybean mutant SS2-2 and to determine whether the gene controlling the hypernodulation of SS2-2 is allelic to that controlling the supernodulation of nts382 mutant. Hybridization studies between SS2-2 and Taekwangkong revealed that the recessive gene was responsible for the hypernodulation character in soybean mutant SS2-2. Allelism was also tested by crossing supernodulating mutant nts382 and hypernodulating mutant SS2-2 that both hypernodulation and supernodulation genes were likely controlled by an identical locus. Molecular marker mapping of hypernodulation gene in SS2-2 using SSR markers confirmed that the gene conferring hypernodulation was located at the same loci with the gene conferring supernodulation. It is interesting to note that the same gene controlled the super- and hyper-nodulation characters, although SS2-2 and nts 382 exhibited differences in the amount of nodulation in the root system. Further genetic studies should be needed to clarify the genetic regulation of super- and hyper-nodulation in soybean.

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Analysis of Korean japonica rice cultivars using molecular markers associated with blast resistance genes

  • Suh, Jung-Pil;Roh, Jae-Hwan;Cho, Young-Chan;Han, Seong-Sook;Jeon, Yong-Hee;Kang, Kyung-Ho;Kim, Yeon-Gyu
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.215-222
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    • 2008
  • Fifty-two Korean japonica rice cultivars were analyzed for leaf blast resistance and genotyped with 4 STS and 26 SSR markers flanking the specific chromosome sites linked with blast resistance genes. In our analysis of resistance genes in 52 japonica cultivars using STS markers tightly linked to Pib, Pita, Pi5(t) and Pi9(t), the blast nursery reaction of the cultivars possessing the each four major genes were not identical to that of the differential lines. Eight of the 26 SSR markers were associated with resistant phenotypes against the isolates of blast nursery as well as the specific Korean blast isolates, 90-008 (KI-1113), 03-177 (KJ-105). These markers were linked to Pit, Pish, Pib, Pi5(t), Piz, Pia, Pik, Pi18, Pita and Pi25(t) resistance gene loci. Three of the eight SSR markers, MRG5836, RM224 and RM7102 only showed significantly associated with the phenotypes of blast nursery test for two consecutive years. These three SSR markers also could distinguish between resistant and susceptible japonica cultivars. These results demonstrate the usefulness of marker-assisted selection and genotypic monitoring for blast resistance of rice in blast breeding programs.

Development of SCAR Marker for Identification of the Perilla Species

  • Lee, Myoung-Hee;Yang, Ki-Woong;Ha, Tae Joung;Jung, Chan-Sik;Pae, Suk-Bok;Hwang, Chung-Dong;Park, Chang-Hwan;Baek, In-Youl;Kim, Hyeon-Kyeong;Park, Soon-Ki
    • Korean Journal of Breeding Science
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    • v.43 no.4
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    • pp.265-272
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    • 2011
  • This study is to generate SCARs markers for identification of Perilla species. A SCAR is a genomic DNA fragment at a single genetically defined locus that is identified by PCR amplification using a pair of specific oligonucleotide primers. We derived SCARs by sequencing and cloning the both ends of the amplified products of RAPD markers. Sixteen sequence-specific primers were synthesized from eight RAPD markers, which were completely sequenced. We developed the species-specific SCAR markers which could be used successfully in detecting genetic variation in four Perilla species. These markers could be used to verify species-origins of various forms of Perilla germplasms.