• Title/Summary/Keyword: Gene Analysis

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Oral Metagenomic Analysis Techniques

  • Chung, Sung-Kyun
    • Journal of dental hygiene science
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    • v.19 no.2
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    • pp.86-95
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    • 2019
  • The modern era of microbial genome analysis began in earnest in the 2000s with the generalization of metagenomics and gene sequencing techniques. Studying complex microbial community such as oral cavity and colon by a pure culture is considerably ineffective in terms of cost and time. Therefore, various techniques for genomic analysis have been developed to overcome the limitation of the culture method and to explore microbial communities existing in the natural environment at the gene level. Among these, DNA fingerprinting analysis and microarray chip have been used extensively; however, the most recent method of analysis is metagenomics. The study summarily examined the overview of metagenomics analysis techniques, as well as domestic and foreign studies on disease genomics and cluster analysis related to oral metagenome. The composition of oral bacteria also varies across different individuals, and it would become possible to analyze what change occurs in the human body depending on the activity of bacteria living in the oral cavity and what causality it has with diseases. Identification, isolation, metabolism, and presence of functional genes of microorganisms are being identified for correlation analysis based on oral microbial genome sequencing. For precise diagnosis and treatment of diseases based on microbiome, greater effort is needed for finding not only the causative microorganisms, but also indicators at gene level. Up to now, oral microbial studies have mostly involved metagenomics, but if metatranscriptomic, metaproteomic, and metabolomic approaches can be taken together for assessment of microbial genes and proteins that are expressed under specific conditions, then doing so can be more helpful for gaining comprehensive understanding.

Genetic Analysis of Wheat for Plant Height by RNA-seq Analysis of Wheat Cultivars 'Keumkang' and 'Komac 5'

  • Moon Seok Kim;Jin Seok Yoon;Yong Weon Seo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.275-275
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    • 2022
  • One of the most widely grown food crops in the world, wheat, is increasing more lodged since for increased rains and winds caused by abnormal climate. During the Green Revolution, shorter wheat cultivars were bred using many Rht genes to increase lodging resistance. However, since only some Rht genes were used for breeding shorter wheat, it may have had a limited impact on wheat breeding and reduced genetic diversity. Therefore, it is essential to search for genes that have breeding potential and affect dwarfism in order to increase the genetic diversity of dwarf characteristics in wheat. In this study, we performed the RNA-seq between 'Keumkang' and 'Komac 5' ('Keumkang' mutant) to analyze the difference in plant height. Differentially expressed genes (DEGs) analysis and Gene function annotation were performed using 265,365,558 mapped reads. Cluster set analysis was performed to compress and select candidate gene DEGs affecting plant height, stem and internode. Gene expression analysis was performed in order to identify the functions of the selected genes by condensing the results of the DEG analysis into a cluster set analysis. This analysis of these plant height-related genes could help reduce plant height, improve lodging resistance, and increase wheat yield. Its application to wheat breeding will also affect the increased genetic diversity of wheat dwarfism.

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Genetic Characteristics of Porcine Epidemic Diarrhea Virus Isolated in Korea (돼지 유행성 설사 바이러스 국내분리주의 유전학적 특성 규명)

  • Chi, Yong-zhe;Kwon, Hyuk-moo;Jeong, Hyun-kyu;Han, Jeong-hee
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.219-230
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    • 2003
  • Porcine epidemic diarrhea virus(PED), a member of Coronaviridea, is the etiological agent of enteropathogenic diarrhea in swine. The purpose of this study was to investigate genetic characteristic of PEDV isolated in Korea. Nucleocapsid(N) gene and membrane (M) gene of recent Korean PEDV strains isolated in 2001 were amplified, cloned, sequenced and analyzed. N gene of seven Korean PEDV field isolates bad 94.5% to 99.4% nucleotide and 92.4% to 99.4% amino acid sequence homology each other. Nucleotide and amino acid sequences of Korean field PEDVs were different from published foreign PEDVs, showing 95.1% to 98.0% nucleotide and 93.5% to 97.6% amino acid sequence homology. By phylogenetic tree analysis on based nucleotide sequences, PEDVs were clustered into four groups. By phylogenetic tree analysis based on amino acid sequences. PEDVs were clustered into five groups. M gene of our Korean PEDV field isolates had 99.6% to 100% nucleotide and 98.7% to 100% amino acid sequence homology each other. Nuclotide and amino acid sequences of Korean field PEDVs were different from published foreign PEDVs, showing 98.5% to 98.8% nucleotide and 97.3% to 97.8% amino acid sequence homology. By phylogenetic tree analysis based on nucleotide and amino acid sequences, PEDVs were clustered into two groups which were Korean PEDV isolate group and foreign PEDV isolate group.

Identification of Gene Expression Signatures in the Chicken Intestinal Intraepithelial Lymphocytes in Response to Herb Additive Supplementations

  • Won, Kyeong-Hye;Song, Ki-Duk;Park, Jong-Eun;Kim, Duk-Kyung;Na, Chong-Sam
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1515-1521
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    • 2016
  • Anethole and garlic have an immune modulatory effects on avian coccidiosis, and these effects are correlated with gene expression changes in intestinal epithelial lymphocytes (IELs). In this study, we integrated gene expression datasets from two independent experiments and investigated gene expression profile changes by anethole and garlic respectively, and identified gene expression signatures, which are common targets of these herbs as they might be used for the evaluation of the effect of plant herbs on immunity toward avian coccidiosis. We identified 4,382 and 371 genes, which were differentially expressed in IELs of chickens supplemented with garlic and anethole respectively. The gene ontology (GO) term of differentially expressed genes (DEGs) from garlic treatment resulted in the biological processes (BPs) related to proteolysis, e.g., "modification-dependent protein catabolic process", "proteolysis involved in cellular protein catabolic process", "cellular protein catabolic process", "protein catabolic process", and "ubiquitin-dependent protein catabolic process". In GO analysis, one BP term, "Proteolysis", was obtained. Among DEGs, 300 genes were differentially regulated in response to both garlic and anethole, and 234 and 59 genes were either up- or down-regulated in supplementation with both herbs. Pathway analysis resulted in enrichment of the pathways related to digestion such as "Starch and sucrose metabolism" and "Insulin signaling pathway". Taken together, the results obtained in the present study could contribute to the effective development of evaluation system of plant herbs based on molecular signatures related with their immunological functions in chicken IELs.

VHL Gene Mutation Analysis of a Chinese Family with Non-Syndromic Pheochromocytomas and Patients with Apparently Sporadic Pheochromocytoma

  • Zhang, Bin;Qian, Jing;Chang, De-Hui;Wang, Yang-Min;Zhou, Da-Hai;Qiao, Gou-Mei
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.1977-1980
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    • 2015
  • Objective: The Von Hippel-Lindau syndrome (VHLD), an inherited neoplastic syndrome predisposing to central nervous system hemangioblastoma (CNS), pheochromocytoma (PCC), renal cell carcinoma(RCC), retinal hemangioma (RA) and renal cysts, is caused by mutations or deletions of the VHL tumor-suppressor gene. To assess VHL genotype-phenotype correlations with function of pVHL a gene mutation analysis of members in a Chinese family with non-syndromic PCCs and individuals with apparently sporadic pheochromocytoma (ASP) was performed. Materials and Methods: DNA samples of 20 members from the Chinese family with non-syndromic PCCs and 41 patients with ASP were analyzed by polymerase chain reaction and direct sequencing, confirmed by Taqman probe. Results: Three novel mutations (H125P, 623(^TTTGTtG) and R120T) were identified in the Chinese family and in 3 among 41 ASP patients. The mutations were all located in exon 2 of VHL gene encoding ${\beta}$-domain of pVHL. The tumor type in H125P carriers and R120T carriers was VHL type 2C. And 623(^TTTGTtG) carriers presented VHL type 2B or type 2C. Conclusions: VHL gene abnormalities were identified in the Chinese family with non-syndromic PCCs and patients with APS, resulting in dysfunction of pVHL. H125P and R120T could be associated with VHL type 2C, while 623(^TTTGTtG) might be linked with VHL type 2B or type 2C. Not only is the genetic analysis helpful for early diagnosis and treatment of patients with VHLD, it is also benefitial for research intoVHLD pathogenesis.

STAT6 Gene Polymorphisms in Allergic Rhinitis

  • Kim, Jeong Joong;Kim, Min Su;Lee, Jung Hun;Choi, Tae Wook;Choi, Sang Heon;Lee, Jae Hoon;Chung, Hun Taeg
    • Genomics & Informatics
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    • v.2 no.3
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    • pp.126-130
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    • 2004
  • T helper-type 2 cytokines, such as IL-4 and IL-13, may play a central role in allergic diseases. The protein known as signal transducers and activators of transcription 6 (STAT6) is a key transcription factor involved in both IL-4- and -13-mediated biological responses. Two polymorphisms of the STAT 6 gene (exon 1 and G2964A variant) have been found. We investigated whether these STAT6 gene polymorph isms were associated with allergic rhinitis. Blood samples for genetic analysis were obtained from 285 individuals with allergic rhinitis and from 271 healthy subjects without atopic disease. The G2964A variant of the STAT6 gene was genotyped using PCR-RFLP analysis. The GT repeat polymorphism in exon 1 of the STAT6 gene was genotyped by fragment analysis. There was no association between the 2964A variant and GT repeat polymorphism in exon 1 of the STAT6 and allergic rhinitis in a Korean population (both p > 0.05). Our results suggest that a combination of STAT6 gene polymorphisms is not a useful marker for predicting allergic rhinitis.

Genetic analysis of clubroot resistance in Chinese cabbage using single spore isolate of Plasmodiophora brassicae and development of RAPD marker linked to its resistance gene

  • Cho, Kwang-Soo;Hong, Su-Young;Han, Young-Han;Yoon, Bong-Kyeong;Ryu, Seoung-Ryeol;Woo, Jong-Gyu
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.101-106
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    • 2008
  • To identify inheritance of clubroot disease resistance genes in Chinese cabbage, seedling tests of $BC_1P_1,\;BC_1P_2$, and $F_2$ populations derived from $F_1$ hybrid(var. CR Saerona) using single spore isolate(race 4 identified with William's differential host) from Plasmodiophora brassciae were conducted. Resistance(R) and susceptible(S) plants segregated to 1:0 in backcross to the resistant parent. The $F_2$ population segregated in a 3(R):1(S) ratio. This result implied that the resistance of clubroot disease is controlled by a single dominant gene to the race 4 of P. brassicae in CR Saerona. To develop DNA markers linked to clubroot resistance genes, 185 plants of CR Saerona among $F_2$ populations were used. A total of 300 arbitrary decamer was applied to $F_2$ population using BSARAPD(Bulked segregant analysis-Randomly amplified polymorphic DNA). One RAPD marker linked to clubroot resistance gene in CR Saerona($OPJ_{1100}$) was identified. This marker was 3.1 cM in distance from resistance gene in $F_2$ population. This marker may be useful for a marker-assisted selection(MAS) and gene pyramiding of the clubroot disease resistant gene in Chinese cabbage breeding programs.

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Development of a Transient ihpRNA-induced Gene Silencing System for Functional Analysis in Persimmon (Diospyros kaki Thunb.)

  • Mo, Rongli;Zhang, Na;Yang, Sichao;Zhang, Qinglin;Luo, Zhengrong
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.314-323
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    • 2016
  • A transient ihpRNA-induced gene silencing system based on Agrobacterium-mediated injection infiltration has been established to evaluate candidate genes involved in proanthocyanidin (PAs) biosynthesis in persimmon (Diospyros kaki Thunb.). We chose DkPDS (phytoene desaturase) as a gene-silencing target to evaluate the newly developed transient gene silencing system. Our qRT-PCR analysis indicated that two ihpRNA constructs (pHG-PDS5' and pHG-PDS3') targeted DkPDS, which also led to significantly reduce expression of DkPDS in 'Mopanshi' persimmon leaves. To further confirm the reliability of the system, we successfully utilized it for DkLAR (leucoanthocyanidin reductase) gene silencing. The expression levels of DkLAR in 'Mopanshi' and 'Eshi 1' leaves were ca. 6-fold and ca. 5-fold lower than those in leaves harboring empty vector (pHG-GFP), respectively. DMACA (4-dimethylaminocinnamaldehyde) staining and the Folin-Ciocalteau assay showed that the accumulation of PAs was markedly inhibited in 'Mopanshi', 'Eshi 1' and 'Youhou' leaves. These results indicate that DkLAR plays an important role in the accumulation of PAs in persimmon. The transient ihpRNA-induced gene silencing method developed in this study is a highly efficient and useful tool for functional analysis of persimmon genes involved in PA biosynthesis.

Characterization and gene expression of heat shock protein 90 in marine crab Charybdis japonica following bisphenol A and 4-nonylphenol exposures

  • Park, Kiyun;Kwak, Ihn-Sil
    • Environmental Analysis Health and Toxicology
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    • v.29
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    • pp.2.1-2.7
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    • 2014
  • Objectives Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone important in the maturation of a broad spectrum of protein. In this study, an HSP90 gene was isolated from Asian paddle crab, Charybdis japonica, as a bio-indicator to monitor the marine ecosystem. Methods This work reports the responses of C. japonica HSP90 mRNA expression to cellular stress by endocrine disrupting chemicals (EDCs), such as bisphenol A (BPA) and 4-nonylphenol (NP) using real-time. reverse transcription polymerase chain reaction. Results The deduced amino acid sequence of HSP90 from C. japonica shared a high degree of homology with their homologues in other species. In a phylogenetic analysis, C. japonica HSP90 is evolutionally related with an ortholog of the other crustacean species. The expression of HSP90 gene was almost distributed in all the examined tissues of the C. japonica crab but expression levels varied among the different body parts of the crabs. We examined HSP90 mRNA expression pattern in C. japonica crabs exposed to EDCs for various exposure times. The expression of HSP90 transcripts was significantly increased in C. japonica crabs exposed to BPA and NP at different concentrations for 12, 24, 48 and 96 hours. The mRNA expression of HSP90 gene was significantly induced in a concentration- and time-dependent manner after BPA or NP exposures for 96 hours. Conclusions Taken together, expression analysis of Asian paddle crab HSP90 gene provided useful molecular information about crab responses in stress conditions and potential ways to monitor the EDCs stressors in marine environments.

Gene Expression Profile and Its Interpretation in Squamous Cell Lung Cancer

  • Park, Dong-Yoon;Kim, Jung-Min;Kim, Ja-Eun;Yoo, Chang-Hyuk;Lee, Han-Yong;Song, Ji-Young;Hwang, Sang-Joon;Yoo, Jae-Cheal;Kim, Sung-Han;Park, Jong-Ho;Yoon, Jeong-Ho
    • Molecular & Cellular Toxicology
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    • v.2 no.4
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    • pp.273-278
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    • 2006
  • 95 squamous cell lung carcinoma samples (normal tissue: 40 samples, tumor: 55 samples) were analyzed with 8 K cDNA microarray. 1-way ANOVA test was employed to select differentially expressed genes in tumor with FDR<0.01. Among the selected 1,655 genes, final 212 genes were chosen according to the expression fold change and used for following analysis. The expression of up-regulated 64 genes was verified with Reverse Transcription PCR and 10 genes were identified as candidates for SCC markers. In our opinion, those candidates can be exploited as diagnostic or therapeutic purposes. Gene Ontology (GO) based analysis was performed using those 212 genes, and following categories were revealed as significant biological processes: Immune response (GO: 0006955), antigen processing (GO: 0030333), inflammatory response (GO: 0006954), Cell adhesion (GO: 0007155), and Epidermis differentiation (GO: 0008544). Gene set enrichment analysis (GSEA) also carried out on overall gene expression profile with 522 functional gene sets. Glycolysis, cell cycle, K-ras and amino acid biosynthesis related gene sets were most distinguished. These results are consistent with the known characteristics of SCC and may be interconnected to rapid cell proliferation. However, the unexpected results from ERK activation in squamous cell carcinoma gripped our attention, and further studies are under progress.