• Title/Summary/Keyword: Gene Screening

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Screening of Differential Promoter Hypermethylated Genes in Primary Oral Squamous Cell Carcinoma

  • Khor, Goot Heah;Froemming, Gabrielle Ruth Anisah;Zain, Rosnah Binti;Abraham, Mannil Thomas;Thong, Kwai Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8957-8961
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    • 2014
  • Background: Promoter hypermethylation leads to altered gene functions and may result in malignant cellular transformation. Thus, identification of biomarkers for hypermethylated genes could be useful for diagnosis, prognosis, and therapeutic treatment of oral squamous cell carcinoma (OSCC). Objectives: To screen hypermethylated genes with a microarray approach and to validate selected hypermethylated genes with the methylation-specific polymerase chain reaction (MSPCR). Materials and Methods: Genome-wide analysis of normal oral mucosa and OSCC tissues was conducted using the Illumina methylation microarray. The specified differential genes were selected and hypermethylation status was further verified with an independent cohort sample of OSCC samples. Candidate genes were screened using microarray assay and run by MSPCR analysis. Results: TP73, PIK3R5, and CELSR3 demonstrated high percentages of differential hypermethylation status. Conclusions: Our microarray screening and MSPCR approaches revealed that the signature candidates of differentially hypermethylated genes may possibly become potential biomarkers which would be useful for diagnostic, prognostic and therapeutic targets of OSCC in the near future.

Screening of Fruiting Body Formation-Specific Genes from the Medicinal Mushroom Cordyceps militaris MET7903 (약용버섯번데기 동충하초 MET7903의 특이적 자실체형성 유전자의 선별)

  • Yun, Bangung;Chung, Ki-Chul
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.145-148
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    • 2004
  • This study was carried out to screen the fruiting body formation-specific genes from the medicinal mushroom Cordyceps militaris. A cDNA synthesized using total RNA from 4 stages of mushroom development, mycelium, primordium, immature fruiting body and mature fruiting body. Differential expression gene screening was performed by DD-PCR(Differential Display Arbitrary Primer PCR) with cDNA, we sequenced partial 6 genes using pGEM cloning vector. The DNA Sequence of the six DD-PCR products derived from differentially expressed genes was compared to that in the GenBank database by using the NCBI BLAST search to identify similarities to known sequences. Sequence analysis showed that six of DD-PCR products have unknown sequence.

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Analysis of Kernel Hardness of Korean Wheat Cultivars

  • Hong, Byung-Hee;Park, Chul-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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    • pp.78-85
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    • 1999
  • To investigate kernel hardness, a compression test which is widely used to measure the hardness of individual kernels as a physical testing method was made simultaneously with the measurement of friabilin (15KDa) which is strongly associated with kernel hardness and was recently developed as a biochemical marker for evaluating kernel hardness in 79 Korean wheat varieties and experimental lines. With the scattered diagram based on the principal component analysis from the parameters of the compression test, 79 Korean wheat varieties were classified into three groups based on the principal component analysis. Since conventional methods required large amount of flour samples for analysis of friabilin due to the relatively small amount of friabilin in wheat kernels, those methods had limitations for quality prediction in wheat breeding programs. An extraction of friabilin from the starch of a single kernel through cesium chloride gradient centrifugation was successful in this experiment. Among 79 Korean wheat varieties and experimental lines 50 lines (63.3%) exhibited a friabilin band and 29 lines (36.7%) did not show a friabilin band. In this study, lines that contained high maximum force and the lower ratio of minimum force to maximum force showed the absence of the friabilin band. Identification of friabilin, which is the product of a major gene, could be applied in the screening procedures of kernel hardness. The single kernel analysis system for friabilin was found to be an easy, simple and effective screening method for early generation materials in a wheat breeding program for quality improvement.

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Screening of Genes Which are Able to Affect Kalanchoe Vegetative Reproduction (Kalanchoe 식물의 영양 번식에 영향을 줄 수 있는 유전자들의 선발)

  • Jung, Yu-Chul;Chung, Young-Jae;Kim, Dong-Giun
    • Journal of Life Science
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    • v.21 no.6
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    • pp.865-874
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    • 2011
  • The genus Bryophyllum is best known for many of its species having the ability to produce plantlets on their leaves. This phenomenon is also known as vegetative reproduction. Differential expressed gene (DEG) detecting technique was applied in order to survey the genes involved in the process of asexual reproduction for plantlet formation. Based on homology search using the NCBI database after screening of genes, 38 genes were identified from a total of 69 DEGs. Most of these DEGs were related to cell division, to intercellular signal transduction, and to hormone (cytokinin and ethylene) signaling.

Screening and Application of Bacillus Strains Isolated from Nonrhizospheric Rice Soil for the Biocontrol of Rice Blast

  • Sha, Yuexia;Zeng, Qingchao;Sui, Shuting
    • The Plant Pathology Journal
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    • v.36 no.3
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    • pp.231-243
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    • 2020
  • Rice blast, caused by Magnaporthe oryzae, is one of the most destructive rice diseases worldwide. The aim of this study was to screen bacterial isolates to efficiently prevent the occurrence of rice blast. A total of 232 bacterial isolates were extracted from nonrhizospheric rice soil and were screened for antifungal activity against M. oryzae using a leaf segment assay. Strains S170 and S9 showed significant antagonistic activity against M. oryzae in vitro and in leaf disk assays, and controlled M. oryzae infection under greenhouse conditions. The results showed that strains S170 and S9 could effectively control rice leaf blast and panicle neck blast after five spray treatments in field. This suggested that the bacterial strains S170 and S9 were valuable and promising for the biocontrol of rice disease caused by M. oryzae. Based on 16S rDNA, and gyrA and gyrB gene sequence analyses, S170 and S9 were identified as Bacillus amyloliquefaciens and B. pumilus, respectively. The research also demonstrated that B. amyloliquefaciens S170 and B. pumilus S9 could colonize rice plants to prevent pathogenic infection and evidently suppressed plant disease caused by 11 other plant pathogenic fungi. This is the first study to demonstrate that B. amyloliquefaciens and B. pumilus isolated from nonrhizospheric rice soil are capable of recolonizing internal rice stem tissues.

Q-omics: Smart Software for Assisting Oncology and Cancer Research

  • Lee, Jieun;Kim, Youngju;Jin, Seonghee;Yoo, Heeseung;Jeong, Sumin;Jeong, Euna;Yoon, Sukjoon
    • Molecules and Cells
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    • v.44 no.11
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    • pp.843-850
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    • 2021
  • The rapid increase in collateral omics and phenotypic data has enabled data-driven studies for the fast discovery of cancer targets and biomarkers. Thus, it is necessary to develop convenient tools for general oncologists and cancer scientists to carry out customized data mining without computational expertise. For this purpose, we developed innovative software that enables user-driven analyses assisted by knowledge-based smart systems. Publicly available data on mutations, gene expression, patient survival, immune score, drug screening and RNAi screening were integrated from the TCGA, GDSC, CCLE, NCI, and DepMap databases. The optimal selection of samples and other filtering options were guided by the smart function of the software for data mining and visualization on Kaplan-Meier plots, box plots and scatter plots of publication quality. We implemented unique algorithms for both data mining and visualization, thus simplifying and accelerating user-driven discovery activities on large multiomics datasets. The present Q-omics software program (v0.95) is available at http://qomics.sookmyung.ac.kr.

Effective Application of Multiplex RT-PCR for Characterization of Human Embryonic Stem Cells/ Induced Pluripotent Stem Cells (다중 역전사 중합효소 연쇄 반응(Multiplex RT-PCR)을 이용한 인간배아 줄기세포 및 유도만능 줄기세포의 효과적인 분화 양상 조사)

  • Kim, Jung-Mo;Cho, Youn-Jeong;Son, On-Ju;Hong, Ki-Sung;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.1-8
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    • 2011
  • Techniques to evaluate gene expression profiling, such as sufficiently sensitive cDNA microarrays or real-time quantitative PCR, are efficient methods for monitoring human pluripotent stem cell (hESC/iPSC) cultures. However, most of these high-throughput tests have a limited use due to high cost, extended turn-around time, and the involvement of highly specialized technical expertise. Hence, there is an urgency of rapid, cost-effective, robust, yet sensitive method development for routine screening of hESCs/hiPSCs. A critical requirement in hESC/hiPSC cultures is to maintain a uniform undifferentiated state and to determine their differentiation capacity by showing the expression of gene markers representing all three germ layers, including ectoderm, mesoderm, and endoderm. To quantify the modulation of gene expression in hESCs/hiPSC during their propagation, expansion, and differentiation via embryoid body (EB) formation, we developed a simple, rapid, inexpensive, and definitive multimarker, semiquantitative multiplex RT-PCR platform technology. Among the 9 gene primers tested, 5 were pluripotent markers comprising set 1, and 3 lineage-specific markers were combined as set 2, respectively. We found that these 2 sets were not only effective in determining the relative differentiation in hESCs/hiPSCs, but were easily reproducible. In this study, we used the hES/hiPS cell lines to standardize the technique. This multiplex RT-PCR assay is flexible and, by selecting appropriate reporter genes, can be designed for characterization of different hESC/hiPSC lines during routine maintenance and directed differentiation.

Characterization of Late-Onset Citrullinemia 1 in a Korean Patient: Confirmation by Argininosuccinate Synthetase Gene Mutation Analysis

  • Kim, In-Suk;Ki, Chang-Seok;Kim, Jong-Won;Lee, Mun-Hyang;Jin, Dong-Kyu;Lee, Soo-Youn
    • BMB Reports
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    • v.39 no.4
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    • pp.400-405
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    • 2006
  • A 16-month old boy was referred to our hospital for evaluation of recurrent generalized tonic clonic seizures. Metabolic evaluation revealed significant hyperammonemia ($1,112\;{\mu}g/dl$). Amino acid/acylcarnitine screening using tandem mass spectrometry showed markedly increased plasma levels of citrulline ($1,350\;{\mu}M/l$) with undetectable levels of arginine and arginosuccinic acid. Urinary excretion of citrulline was markedly increased ($38,617\;{\mu}M/g$ creatinine). Brain MRI findings showed diffuse high-signal intensity lesions, that involved gray and white matter in both frontal lobes and insula with edematous changes; these findings were consistent with the acute stage of citrullinemia (CTLN). Mutation analysis of the argininosuccinate synthetase (ASS) gene, in this patient, showed a Gly324Ser mutation in exon 13, and a 67-bp duplication mutation in exon 15 (c.1128-6_1188dup67). The patient was confirmed as having late-onset CTLN1 and treated with anticonvulsants, lactulose enema, protein restricted diet and arginine. Here we describe a case of late-onset CTLN1 in a patient by biochemical analyses and ASS gene mutation confirmation. This is the first report of a Korean patient with late-onset CTLN1 confirmed by ASS gene mutation identification.

Screening of the Dominant Rice Blast Resistance Genes with PCR-based SNP and CAPS Marker in Aromatic Rice Germplasm

  • Kim, Jeong-Soon;Ahn, Sang-Nag;Hong, Sung-Jun;Kwon, Jin-Hyeuk;Kim, Yeong-Ki;Jee, Hyeong-Jin;Shim, Chang-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.4
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    • pp.329-341
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    • 2011
  • The objective of this study was to determine the genetic diversities of major rice blast resistance genes among 84 accessions of aromatic rice germplasm. Eighty four accessions were characterized by a dominant 11 set of PCR-based SNP and CAPS marker, which showed the broad spectrum resistance and closest linkage to seven major rice blast resistance (R) genes, Pia, Pib, Pii, Pi5 (Pi3), Pita (Pita-2), and Pi9 (t). The allele specific PCR markers assay genotype of SCAR and STS markers was applied to estimate the presence or absence of PCR amplicons detected with a pair of PCR markers. One indica accession, Basmati (IT211194), showed the positive amplicons of five major rice blast resistance genes, Pia, Pi5 (Pi3), Pib, Pi-ta (Pi-ta2), and Pik-5 (Pish). Among 48 accessions of the PCR amplicons detected with yca72 marker, only five accessions were identified to Pia gene on chromosome 11. The Pib gene was estimated with the NSb marker and was detected in 65 of 84 accessions. This study showed that nine of 84 accessions contained the Pii gene and owned Pi5 (Pi3) in 42 of 84 accessions by JJ817 and JJ113-T markers, which is coclosest with Pii on chromosome 9. Only six accessions were detected two alleles of the Pita or Pita-2 genes. Three of accessions were identified as the Pi9 (t) gene locus.

Gene Cloning and Nucleotide Sequence of Human Dihydrolipoamide Dehydrogenase-Binding Protein

  • Lee, Jeongmin;Ryou, Chongsuk;Jeon, Bong Kyun;Lee, Poongyeon;Woo, Hee-Jong;Kwon, Moosik
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.3
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    • pp.421-426
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    • 2002
  • The pyruvate dehydrogenase complex (PDC), a member of $\alpha$-keto acid dehydrogenase complex, catalyzes the oxidative decarboxylation of pyruvate with the formation of $CO_2$, acetyl-CoA, NADH, and $H^+$. This complex contains multiple copies of three catalytic components including pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide dehydrogenase (E3). Two regulatory components (E1-kinase and phospho-E1 phosphatase) and functionally less-understood protein (protein X, E3BP) are also involved in the formation of the complex. In this study, we have partially cloned the gene for E3BP in human. Nine putative clones were isolated by human genomic library screening with 1.35 kb fragment of E3BP cDNA as a probe. For investigation of cloned genes, Southern blot analysis and the construction of the restriction map were performed. One of the isolated clones, E3BP741, has a 3 kb-SacI fragment, which contains 200 bp region matched with E3BP cDNA sequences. The matched DNA sequence encodes the carboxyl-terminal portion of lipoyl-bearing domain and hinge region of human E3BP. Differences between yeast E3BP and mammalian E3BP coupled with the remarkable similarity between mammalian E2 and mammalian E3BP were confirmed from the comparison of the nucleotide sequence and the deduced amino acid sequence in the cloned E3BP. Cloning of human E3BP gene and analysis of the gene structure will facilitate the understanding of the role(s) of E3BP in mammalian PDC.