• Title/Summary/Keyword: multiple genes

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Analysis of 16S-23S rRNA Intergenic Spacer Region of Vibrio vulnificus (Vibrio vulnificus의 16S-23S rRNA Intergenic Spacer Region 분석)

  • PARK Young Mi;LEE Jehee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.3
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    • pp.239-246
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer region (ISR) of Vibrio vulnificus KCTC 2959. ISRs were amplified by primers complementary to conserved regions of 16S and 23S rRNA genes. ISR amplicons were cloned and sequenced. Analysis of the ISR sequences showed that V. vulnificus KCTC 2959 contains five types of polymorphic ISRs. Size of ISRs ranged from 424 to 741 bp in length and the number of tRNA genes ranged from one to four. The ISRs were designated as ISR-E $(tRNA^{Glu}),\;ISR-IA\;(tRNA^{Ile}-tRNA^{Ala})$, ISR-EKV $(tRNA^{Glu}-tRNA^{Lys}-tRNA^{Val})$, ISR-IAV $(tRNA^{Ile}-tRNA^{Ala}-tRNA^{val})$ and ISR-EKAV $(tRNA^{Glu}-tRNA^{Lys}-tRNA^{Ala}-tRNA^{Val})$ based on their tRNA genes. Multiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability. We used the sequences of variable domains to design species-specific primer for detection PCR. Specificity of the primers was examined using genomic DNA prepared from 18 different Vibrio species. The results showed that the PCR using primers designed in this study can be used to detect V. vulnificus from other Vibrio species.

Integrative Meta-Analysis of Multiple Gene Expression Profiles in Acquired Gemcitabine-Resistant Cancer Cell Lines to Identify Novel Therapeutic Biomarkers

  • Lee, Young Seok;Kim, Jin Ki;Ryu, Seoung Won;Bae, Se Jong;Kwon, Kang;Noh, Yun Hee;Kim, Sung Young
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.2793-2800
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    • 2015
  • In molecular-targeted cancer therapy, acquired resistance to gemcitabine is a major clinical problem that reduces its effectiveness, resulting in recurrence and metastasis of cancers. In spite of great efforts to reveal the overall mechanism of acquired gemcitabine resistance, no definitive genetic factors have been identified that are absolutely responsible for the resistance process. Therefore, we performed a cross-platform meta-analysis of three publically available microarray datasets for cancer cell lines with acquired gemcitabine resistance, using the R-based RankProd algorithm, and were able to identify a total of 158 differentially expressed genes (DEGs; 76 up- and 82 down-regulated) that are potentially involved in acquired resistance to gemcitabine. Indeed, the top 20 up- and down-regulated DEGs are largely associated with a common process of carcinogenesis in many cells. For the top 50 up- and down-regulated DEGs, we conducted integrated analyses of a gene regulatory network, a gene co-expression network, and a protein-protein interaction network. The identified DEGs were functionally enriched via Gene Ontology hierarchy and Kyoto Encyclopedia of Genes and Genomes pathway analyses. By systemic combinational analysis of the three molecular networks, we could condense the total number of DEGs to final seven genes. Notably, GJA1, LEF1, and CCND2 were contained within the lists of the top 20 up- or down-regulated DEGs. Our study represents a comprehensive overview of the gene expression patterns associated with acquired gemcitabine resistance and theoretical support for further clinical therapeutic studies.

Alleviation of Salt Stress by Enterobacter sp. EJ01 in Tomato and Arabidopsis Is Accompanied by Up-Regulation of Conserved Salinity Responsive Factors in Plants

  • Kim, Kangmin;Jang, Ye-Jin;Lee, Sang-Myeong;Oh, Byung-Taek;Chae, Jong-Chan;Lee, Kui-Jae
    • Molecules and Cells
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    • v.37 no.2
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    • pp.109-117
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    • 2014
  • Microbiota in the niches of the rhizosphere zones can affect plant growth and responses to environmental stress conditions via mutualistic interactions with host plants. Specifically, some beneficial bacteria, collectively referred to as Plant Growth Promoting Rhizobacteria (PGPRs), increase plant biomass and innate immunity potential. Here, we report that Enterobacter sp. EJ01, a bacterium isolated from sea china pink (Dianthus japonicus thunb) in reclaimed land of Gyehwa-do in Korea, improved the vegetative growth and alleviated salt stress in tomato and Arabidopsis. EJ01 was capable of producing 1-aminocy-clopropane-1-carboxylate (ACC) deaminase and also exhibited indole-3-acetic acid (IAA) production. The isolate EJ01 conferred increases in fresh weight, dry weight, and plant height of tomato and Arabidopsis under both normal and high salinity conditions. At the molecular level, short-term treatment with EJ01 increased the expression of salt stress responsive genes such as DREB2b, RD29A, RD29B, and RAB18 in Arabidopsis. The expression of proline biosynthetic genes (i.e. P5CS1 and P5CS2) and of genes related to priming processes (i.e. MPK3 and MPK6) were also up-regulated. In addition, reactive oxygen species scavenging activities were enhanced in tomatoes treated with EJ01 in stressed conditions. GFP-tagged EJ01 displayed colonization in the rhizosphere and endosphere in the roots of Arabidopsis. In conclusion, the newly isolated Enterobacter sp. EJ01 is a likely PGPR and alleviates salt stress in host plants through multiple mechanisms, including the rapid up-regulation of conserved plant salt stress responsive signaling pathways.

Genetic interaction of Sub1A and Pup1 in rice

  • Shin, Na-Hyun;Yoo, Soo-Cheul;Chin, Joong Hyoun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.147-147
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    • 2017
  • Rice is one of the major staple food in Asia, covering around half of the world population. More than 40% of rice cultivation area are subject to abiotic stresses such as drought, submergence and phosphate deficiency. Pyramiding useful genes into elite variety is a promising strategy to develop tolerance varieties to multiple abiotic stresses. However, some genes are not functionally compatible when they are introgressed into the same elite variety. Here, we tested the functional compatibility of Sub1 and Pup1, major QTLs for tolerance to submergence and phosphate (P)-deficiency conditions, respectively. Phenotypic analysis revealed that IR64-Sub1 Pup1(SP1) plants harboring both Sub1 and Pup1 QTLs showed significant tolerance to submerged conditions, similarly in IR64-Sub1 (Sub1) plant, while SP1 plants failed to tolerate to P-deficiency conditions; only IR64-Pup1 (Pup1) showed strong P-deficiency tolerance phenotype. In submerged conditions, the expression levels of Sub1A and PSTOL1, major genes for Sub1 and Pup1 QTLs, respectively, were not significantly different in between Pup1 and SP1 plants. On the other hand, the expression of both Sup1A and PSTOL1 was significantly downregulated in P-deficiency conditions, suggesting that Sub1 and Pup1 repressed gene expression each other in P-deficiency conditions. These results suggest Pup1 does not compromise the Sub1 function in submerged conditions while Sub1 suppresses the function of Pup1 in (P)-deficient condition, possibly by regulating transcript level of Pup1. In conclusion, Sub1 and Pup1 are functionally compatible in terms of submergence tolerance but not in P-deficiency conditions. Further analysis need to be performed to elucidate how Sup1 suppresses the function of Pup1 in P-deficiency conditions.

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Characterization of Extended-Spectrum-$\beta$-Lactamase Genotype TEM, SHV and CTX-M from Clinical Isolates of Klebsiella pneumoniae and Comparison with Antibiotic Susceptibility Test

  • Kim Yun-Tae;Oh Kwang-Seok;Choi Seok-Cheol;Kim Tae-Un
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.389-396
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    • 2005
  • Resent studies have reported increased isolation of extended-spectrum $\beta-lactamase$ (ESBL) producing strains at several hospital in Korea. We studied to investigate the isolation rates of ESBL strains from clinical isolates of Klebsiella pneumoniae and to characterize differences in types using analyses of genotyping and antibiotic susceptibility test. Antibiotic susceptibility test with confirmation of ESBL by double disk synergy test was performed on the 54 ESBL strains of Klebsiella pneumoniae from a hospital in Busan. Transfer of resistant gene in ESBL strains resistant to 3rd generated antibiotics was confirmed by transconjugation test using E. coli $RG176^{nal(r)}$. blaTEM, blaSHV, blaCTX-M genes were detected by PCR. ESBL producing strains had 100% of resistant rate to ampicillin, azteronam, cefazolin, cefepime and ceftriaxone ($\beta-lactam$ antibiotics). Forty strains of bla TEM$(74\%)$, 41 strains of bla SHV $(76\%)$, 23 strains of bla CTX-M $(43\%)$ were found, respectively. The strains had one or more genes. They had high resistant rates to $\beta-lactam$ antibiotics including cephalosporin. The resistant rates of strains with multiple resistant genes were higher than those of strains with single resistant gene.

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Recent Molecular Studies of Marek's Disease Virus and Control Approaches for Increasing Virulence (마렉병 바이러스의 분자적 병리기전과 증가하는 병원성에 대한 제어 방안)

  • Jang, H.K.;Park, Y.M.;Cha, S.Y.;Kwon, J.T.
    • Korean Journal of Poultry Science
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    • v.34 no.1
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    • pp.57-76
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    • 2007
  • Marek's disease (MD) is caused by a ubiquitous, lymphotropic alphaherpesvirus, MD virus (MDV). MD has been a major concern in the poultry industry due to the emergence of increasingly virulent strains over the last few decades that were isolated in the face of comprehensive vaccination. MD is characterized by a variety of clinical signs, amongst them neurological symptoms, chronic wasting, and most notably the development of multiple lymphomas that manifest as solid tumors in the viscera and musculature. Much work has been devoted to study MD-induced oncogenesis and genes involved in this process. Among the many genes encoded by MDV, a number have recently been shown to affect the development of tumors in chickens, one protein directly causing transformation of cells (Meq) and another being involved in maintaining transformed cells (vTR). Other MDV gene products modulate and are involved in early lytic in vivo replication, thereby increasing the chance of transformation occurring. In this review, specific genes encoded by MDV that are involved in the initiation and/or maintenance of transformation were briefly summarized, and limits of current vaccination and new control strategies against MD, particularly how modem molecular biological methods may be used to improve strategies to combat the disease in the future, were discussed.

Brassica rapa Sec14-like protein gene BrPATL4 determines the genetic architecture of seed size and shape

  • Kim, Joonki;Lee, Hye-Jung;Nogoy, Franz Marielle;Yu, Dal-A;Kim, Me-Sun;Kang, Kwon-Kyoo;Nou, Illsup;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.332-340
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    • 2016
  • Seed size traits are controlled by multiple genes in crops and determine grain yield, quality and appearance. However, the molecular mechanisms controlling the size of plant seeds remain unclear. We performed functional analysis of BrPATL4 encoding Sec14-like protein to determine the genetic architecture of seed size, shape and their association analyses. We used 60 $T_3$ transgenic rice lines to evaluate seed length, seed width and seed height as seed size traits, and the ratios of these values as seed shape traits. Pleiotropic effects on general architecture included small seed size, erect panicles, decreased grain weight, reduced plant height and increased sterility, which are common to other mutants deficient in gibberellic acid (GA) biosynthesis. To test whether BrPATL4 overexpression is deleterious for GA signal transduction, we compared the relative expression of GA related gene and the growth rate of second leaf sheath supplied with exogenous $GA_3$. Overexpression of BrPATL4 did not affect GA biosynthesis or signaling pathway, with the same response shown under GA treatment compared to the wild type. However, the causal genes for the small seed phenotype (D1, SRS1, and SRS5) and the erection of panicles showed significantly decreased levels in mRNA accumulation compared to the wild type. These results suggest that the overexpression of BrPATL4 can control seed size through the suppression of those genes related to seed size regulation. Although the molecular function of BrPATL4 is not clear for small seed and erect panicles of BrPALT4 overexpression line, this study provides some clues about the genetic engineering of rice seed architecture.

Role of Oxidative Stress and Mitochondria in Parkinson's Disease

  • Jin, Son-Hyeung
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2007.04a
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    • pp.147-153
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    • 2007
  • Central to developing new treatment strategies for late onset sporadic Parkinson's disease (PD) and early onset familial PD is resolving the enigma of the specific vulnerability exhibited by substantia nigra dopamine (DA) neurons despite multiple risk factors. Neuropathological evidence from both human and experimental models of PD firmly supports a significant role for oxidative stress (OS) and mitochondrial dysfunction in the death of nigral DA neurons. Largely unknown are the genes underlying selective susceptibility of nigral DA neuron to OS and mitochondrial dysfunction and how they effect nigral DA cell death. To overcome the paucity of nigral DA neurons as well as the dilution effect of non-DA cells in brain tissues, we have developed wild type DA cell line model, SN4741 and mutant DJ-1 (-/-) DA cells, appropriate for microarray analysis and differential mitochondrial proteomics. Mutations in the DJ-1 gene (PARK7), localized in cytoplasm and mitochondria, cause autosomal recessive early onset PD. Through microarray analysis using SN4741 cells followed by validation tests, we have identified a novel phylogenically conserved neuroprotective gene, Oxi-a, which is specifically expressed in DA neurons. The knockdown of the gene dramatically increased vulnerability to as. Importantly as down-regulated the expression level of the gene and recovery of its expression via transient transfection exerted significant neuroprotection against as insult. We also have identified altered expression of mitochondrial proteins and other familial PD genes in DJ-1 (-/-) mutant cells by differential mitochondrial proteomics. In DJ-1 (-/-) cells the knockdown of the other familial PD genes (Parkin and PINK1) dramatically increased susceptibility to as. Thus, further functional characterization of the Oxi-$\alpha$ gene family and the mitochondrial alteration in the DJ-1 (-/-) cell model will provide the rationale for the neuroprotective therapy against both sporadic and familial PD.

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Analysis of Saccharomyces Cell Cycle Expression Data using Bayesian Validation of Fuzzy Clustering (퍼지 클러스터링의 베이지안 검증 방법을 이용한 발아효모 세포주기 발현 데이타의 분석)

  • Yoo Si-Ho;Won Hong-Hee;Cho Sung-Bae
    • Journal of KIISE:Software and Applications
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    • v.31 no.12
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    • pp.1591-1601
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    • 2004
  • Clustering, a technique for the analysis of the genes, organizes the patterns into groups by the similarity of the dataset and has been used for identifying the functions of the genes in the cluster or analyzing the functions of unknown gones. Since the genes usually belong to multiple functional families, fuzzy clustering methods are more appropriate than the conventional hard clustering methods which assign a sample to a group. In this paper, a Bayesian validation method is proposed to evaluate the fuzzy partitions effectively. Bayesian validation method is a probability-based approach, selecting a fuzzy partition with the largest posterior probability given the dataset. At first, the proposed Bayesian validation method is compared to the 4 representative conventional fuzzy cluster validity measures in 4 well-known datasets where foray c-means algorithm is used. Then, we have analyzed the results of Saccharomyces cell cycle expression data evaluated by the proposed method.

Clinical significance of APOB inactivation in hepatocellular carcinoma

  • Lee, Gena;Jeong, Yun Seong;Kim, Do Won;Kwak, Min Jun;Koh, Jiwon;Joo, Eun Wook;Lee, Ju-Seog;Kah, Susie;Sim, Yeong-Eun;Yim, Sun Young
    • Experimental and Molecular Medicine
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    • v.50 no.11
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    • pp.7.1-7.12
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    • 2018
  • Recent findings from The Cancer Genome Atlas project have provided a comprehensive map of genomic alterations that occur in hepatocellular carcinoma (HCC), including unexpected mutations in apolipoprotein B (APOB). We aimed to determine the clinical significance of this non-oncogenetic mutation in HCC. An Apob gene signature was derived from genes that differed between control mice and mice treated with siRNA specific for Apob (1.5-fold difference; P < 0.005). Human gene expression data were collected from four independent HCC cohorts (n = 941). A prediction model was constructed using Bayesian compound covariate prediction, and the robustness of the APOB gene signature was validated in HCC cohorts. The correlation of the APOB signature with previously validated gene signatures was performed, and network analysis was conducted using ingenuity pathway analysis. APOB inactivation was associated with poor prognosis when the APOB gene signature was applied in all human HCC cohorts. Poor prognosis with APOB inactivation was consistently observed through cross-validation with previously reported gene signatures (NCIP A, HS, high-recurrence SNUR, and high RS subtypes). Knowledge-based gene network analysis using genes that differed between low-APOB and high-APOB groups in all four cohorts revealed that low-APOB activity was associated with upregulation of oncogenic and metastatic regulators, such as HGF, MTIF, ERBB2, FOXM1, and CD44, and inhibition of tumor suppressors, such as TP53 and PTEN. In conclusion, APOB inactivation is associated with poor outcome in patients with HCC, and APOB may play a role in regulating multiple genes involved in HCC development.