• Title/Summary/Keyword: Resistant Genes

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Establishment and Characterization of Carboplatin-Resistant Retinoblastoma Cell Line

  • Cho, Chang Sik;Jo, Dong Hyun;Kim, Jin Hyoung;Kim, Jeong Hun
    • Molecules and Cells
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    • v.45 no.10
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    • pp.729-737
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    • 2022
  • Carboplatin-based chemotherapy is the primary treatment option for the management of retinoblastoma, an intraocular malignant tumor observed in children. The aim of the present study was to establish carboplatin-resistant retinoblastoma cell lines to facilitate future research into the treatment of chemoresistant retinoblastoma. In total, two retinoblastoma cell lines, Y79 and SNUOT-Rb1, were treated with increasing concentrations of carboplatin to develop the carboplatin-resistant retinoblastoma cell lines (termed Y79/CBP and SNUOT-Rb1/CBP, respectively). To verify resistance to carboplatin, the degree of DNA fragmentation and the expression level of cleaved caspase-3 were evaluated in the cells, following carboplatin treatment. In addition, the newly developed carboplatin-resistant retinoblastoma cells formed in vivo intraocular tumors more effectively than their parental cells, even after the intravitreal injection of carboplatin. Interestingly, the proportion of cells in the G0/G1 phase was higher in Y79/CBP and SNUOT-Rb1/CBP cells than in their respective parental cells. In line with these data, the expression levels of cyclin D1 and cyclin D3 were decreased, whereas p18 and p27 expression was increased in the carboplatin-resistant cells. In addition, the expression levels of genes associated with multidrug resistance were increased. Thus, these carboplatin-resistant cell lines may serve as a useful tool in the study of chemoresistance in retinoblastoma and for the development potential therapeutics.

Evaluation of Bacterial Blight Resistance Using SNP and STS Marker-assisted Selection in Aromatic Rice Germplasm

  • Kim, Jeong-Soon;Gwang, Jae-Gyun;Park, Ki-Hun;Shim, Chang-Ki
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.408-416
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    • 2009
  • A molecular survey was conducted to identify the presence of the bacterial blight resistance genes (Xa1, Xa4, xa5, xa13 and Xa21) in 86 accessions of aromatic rice obtained from germplasm. The results revealed that the resistance gene Xa4 (32.5%), Xa21 (17%), and xa5 (16%) were widely observed in tested rice germplasm. Among tested rice germplasm, 49 accessions showed the presence of more than one of five R genes, and 37 accessions possessed none of the R gene. TALLi and 05-IRRi-M-46 showed the presence of Xa4, xa5, xa13 and Xa21. Rice race $415{\times}Ir352$ exhibited positive amplicon for the Xa1, Xa4, xa5 and Xa21. Hyangmibyeo1hos, Ir841-85-1-1-2 and Jasmine85 showed the positive amplicon for the Xa1, Xa4 and xa5 genes. Yekywin Yinkya Hmwe and Khao Dawk Mali105 showed the presence of Xa1, Xa4 and Xa21 gene. Masino Basmati showed the presence of xa5, xa13, Xa21 genes. Xa1 and Xa21 genes were noticed in Mihayngbyeo, Tarana Deshi, Mayataung and AZUCENA. Hyangmibyeo2ho, Basmati 6311 and Basmati405 possessed only two R genes such as Xa4 and xa5, and xa5 and xa13, respectively. The evaluation results of bacterial blight resistance genes in aromatic rice germplasm will help in breeding of multi disease resistant varieties.

Transcription Analysis of Daptomyc in Biosynthetic Genesin Streptomyces roseosporus

  • Rhee, Ki-Hyeong;Davies, Julian
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1841-1848
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    • 2006
  • Insights into gene expression have the potential for improvement of antibiotic yield and the development of robust production hosts for use in recombinant biomolecule production. $Cubicin^{TM}$ (daptomycin for injection) is a recently approved antibiotic active against many Gram(+) pathogens, including those resistant to methicillin, vancomycin, and fluoroquinolones. Daptomycin is produced as a secondary metabolite by Streptomyces roseosporus. A 128 kb region of DNA including the daptomycin biosynthetic gene cluster (dpt) has been cloned. and sequenced. Using a selected array of nucleic acid probes representing this region, we compared the expression levels of the dpt genes between S. roseosporus wild-type (WT) and derived S. roseosporus high-producer of daptomycin (HP). We observed that the majority of the biosynthetic genes were upregulated in HP compared with WT; a total of 12 genes, including those encoding daptomycin synthetase, showed consistently and significantly higher expression levels, at least 5-fold, in HP compared with WT. In contrast, some genes, flanking the dpt cluster, were expressed at higher levels in the WT strain. The expression of housekeeping genes such as S. roseosporus rpsL, rpsG, and 16S (positive controls) and presumptive intergenic regions in the dpt cluster (negative control) were identical in the two strains. In addition, we compared transcription during the early, mid-log, and early-stationary phases of growth in the HP strain. The same set of genes was upregulated and downregulated under all conditions examined; housekeeping genes showed no relative change in expression level over the periods of growth tested. Analyses of this type would be of value in studies of strain improvement and also for the identification of gene regulation processes that are important for secondary metabolite production.

Differential Expressions of Apoptosis-related Genes in Lung Cancer Cell Lines Determine the Responsiveness to Ionizing Radiation

  • Lee, Su-Yeon;Choi, Moon-Kyung;Lim, Jung-Min;Wu, Hong-Gyun;Kim, Ju-Han;Park, Woong-Yang
    • Genomics & Informatics
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    • v.6 no.1
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    • pp.36-43
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    • 2008
  • Radiotherapy would be the choice of treatment for human cancers, because of high cost-effectiveness. However, a certain population of patients shows a resistance to radiotherapy and recurrence. In an effort to increase the efficacy of radiotherapy, many efforts were driven to find the genes causing the unresponsiveness to ionizing radiation. In this paper, we compared the gene expression profiles of two lung cancer cell lines, H460 and H1299, which showed differential responses to ionizing radiations. Each cell were irradiated at 2 Gy, and harvested after 0, 2, 4, 8, 12 and 24 hours to examine the expressions. Two-way ANOVA analysis on time-series experiments of two cells could select 2863 genes differentially expressed upon ionizing radiation among 32,321 genes in microarray (p<0.05). We classified these genes into 21 clusters by SOM clustering according to the interaction between cell types and time. Two SOM clusters were enriched with apoptosis-related genes in pathway analysis. One cluster contained higher levels of phosphatidyl inositol 3-phosphate kinase (PI3K) subunits in H1299, radio-resistant cells than H460, radiosensitive cells. TRAIL receptors were expressed in H460 cells while the decoy receptor for TRAIL was expressed in H1299 cells. From these results, we could characterize the differential responsiveness to ionizing radiation according to their differential expressions of apoptosis-related genes, which might be the candidates to increase the power of radiotherapy.

Transcriptome analyses of the ginseng root rot pathogens Cylindrocarpon destructans and Fusarium solani to identify radicicol resistance mechanisms

  • Li, Taiying;Kim, Jin-Hyun;Jung, Boknam;Ji, Sungyeon;Seo, Mun Won;Han, You Kyoung;Lee, Sung Woo;Bae, Yeoung Seuk;Choi, Hong-Gyu;Lee, Seung-Ho;Lee, Jungkwan
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.161-167
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    • 2020
  • Background: The ascomycete fungi Cylindrocarpon destructans (Cd) and Fusarium solani (Fs) cause ginseng root rot and significantly reduce the quality and yield of ginseng. Cd produces the secondary metabolite radicicol, which targets the molecular chaperone Hsp90. Fs is resistant to radicicol, whereas other fungal genera associated with ginseng disease are sensitive to it. Radicicol resistance mechanisms have not yet been elucidated. Methods: Transcriptome analyses of Fs and Cd mycelia treated with or without radicicol were conducted using RNA-seq. All of the differentially expressed genes (DEGs) were functionally annotated using the Fusarium graminearum transcript database. In addition, deletions of two transporter genes identified by RNA-seq were created to confirm their contributions to radicicol resistance. Results: Treatment with radicicol resulted in upregulation of chitin synthase and cell wall integrity genes in Fs and upregulation of nicotinamide adenine dinucleotide dehydrogenase and sugar transporter genes in Cd. Genes encoding an ATP-binding cassette transporter, an aflatoxin efflux pump, ammonium permease 1 (mep1), and nitrilase were differentially expressed in both Fs and Cd. Among these four genes, only the ABC transporter was upregulated in both Fs and Cd. The aflatoxin efflux pump and mep1 were upregulated in Cd, but downregulated in Fs, whereas nitrilase was downregulated in both Fs and Cd. Conclusion: The transcriptome analyses suggested radicicol resistance pathways, and deletions of the transporter genes indicated that they contribute to radicicol resistance.

Identification of ABSCISIC ACID (ABA) signaling related genes in Panax ginseng

  • Hong, Jeongeui;Kim, Hogyum;Ryu, Hojin
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.306-314
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    • 2018
  • Korean ginseng (Panax ginseng) has long been cultivated as an important economic medicinal plant. Owing to the seasonal and long-term agricultural cultivation methods of Korean ginseng, they are always vulnerable to various environmental stress conditions. ABSCISIC ACID (ABA) is an essential plant hormone associated with seed development and diverse abiotic stress responses including drought, cold and salinity stress. By modulating ABA responses, plants can regulate their immune responses and growth patterns to increase their ability to tolerate stress. With recent advances in genome sequencing technology, we first reported the functional features of genes related to canonical ABA signaling pathway in P. ginseng genome. Based on the protein sequences and functional genomic analysis of Arabidopsis thaliana, the ABA related genes were successfully identified. Our functional genomic characterizations clearly showed that the ABA signaling related genes consisting the ABA receptor proteins (PgPYLs), kinase family (PgSnRKs) and transcription factors (PgABFs, PgABI3s and PgABI5s) were evolutionary conserved in the P. ginseng genome. We confirmed that overexpressing ABA related genes of P. ginseng completely restored the ABA responses and stress tolerance in ABA defective Arabidopsis mutants. Finally, tissue and age specific spatio-temporal expression patterns of the identified ABA-related genes in P. ginseng tissues were also classified using various available RNA sequencing data. This study provides ABA signal transduction related genes and their functional genomic information related to the growth and development of Korean ginseng. Additionally, the results of this study could be useful in the breeding or artificial selection of ginseng which is resistant to various stresses.

Characterization of plasmid-mediated quinolone resistance genes in Enterobacteriaceae isolated from companion animals (반려동물 유래 장내세균에서 plasmid 매개 퀴놀론 내성 유전자의 특성)

  • Cho, Jae-Keun;Kim, Jeong-Mi;Kim, Hwan-Deuk;Kim, Kyung-Hee;Lim, Hyun-Suk;Yang, Chang-Ryoul
    • Korean Journal of Veterinary Service
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    • v.42 no.1
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    • pp.17-24
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    • 2019
  • The aim of this study was to investigate the prevalence and characterization of plasmid-mediated quinolone resistance (PMQR) gene in 79 Enterobacteriaceae isolated from dogs and cats. Of 79 isolates, PMQR genes were found in 10 (12.7%) isolates, including aac(6')-lb-cr, qnrB, qnrS and qnrA detected alone or in combination in 8 (10.1%), 4 (5.1%), 2 (2.5%) and 1 (1.3%) isolates, respectively. Interestingly, two qnrS genes were detected in nalidixic acid and ciprofloxacin susceptible isolates. Extended-spectrum ${\beta}$-lactamase (ESBL) was detected in 90% (9 isolates) of PMQR positives isolates. Among ESBL genes, CTX-M, TEM and SHV were detected in 9, 8 and 3 isolates, respectively. Almost all PMQR genes were detected in co-existence with ESBL genes. All PMQR positives isolates were multidrug resistance (i.e. resistant to five or more antibiotics). qepA, OXA and CMY-2 genes were not found. The six transconjugants were obtained by conjugation experiment. The aac(6')-lb-cr, qnrB and qnrS were co-transferred with CTX-M, TEM and/or SHV, whereas qnrA was not observed among transconugants. This is the first report of the presence of aac(6')-lb-cr and qnrA gene among Enterobacteriaceae isolates from dogs in Korea. The prudent use of antimicrobials and continuous monitoring for companion animals are required.

Expression profiling of cultured podocytes exposed to nephrotic plasma reveals intrinsic molecular signatures of nephrotic syndrome

  • Panigrahi, Stuti;Pardeshi, Varsha Chhotusing;Chandrasekaran, Karthikeyan;Neelakandan, Karthik;PS, Hari;Vasudevan, Anil
    • Clinical and Experimental Pediatrics
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    • v.64 no.7
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    • pp.355-363
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    • 2021
  • Background: Nephrotic syndrome (NS) is a common renal disorder in children attributed to podocyte injury. However, children with the same diagnosis have markedly variable treatment responses, clinical courses, and outcomes, suggesting molecular heterogeneity. Purpose: This study aimed to explore the molecular responses of podocytes to nephrotic plasma to identify specific genes and signaling pathways differentiating various clinical NS groups as well as biological processes that drive injury in normal podocytes. Methods: Transcriptome profiles from immortalized human podocyte cell line exposed to the plasma of 8 subjects (steroid-sensitive nephrotic syndrome [SSNS], n=4; steroid-resistant nephrotic syndrome [SRNS], n=2; and healthy adult individuals [control], n=2) were generated using microarray analysis. Results: Unsupervised hierarchical clustering of global gene expression data was broadly correlated with the clinical classification of NS. Differential gene expression (DGE) analysis of diseased groups (SSNS or SRNS) versus healthy controls identified 105 genes (58 up-regulated, 47 down-regulated) in SSNS and 139 genes (78 up-regulated, 61 down-regulated) in SRNS with 55 common to SSNS and SRNS, while the rest were unique (50 in SSNS, 84 genes in SRNS). Pathway analysis of the significant (P≤0.05, -1≤ log2 FC ≥1) differentially expressed genes identified the transforming growth factor-β and Janus kinase-signal transducer and activator of transcription pathways to be involved in both SSNS and SRNS. DGE analysis of SSNS versus SRNS identified 2,350 genes with values of P≤0.05, and a heatmap of corresponding expression values of these genes in each subject showed clear differences in SSNS and SRNS. Conclusion: Our study observations indicate that, although podocyte injury follows similar pathways in different clinical subgroups, the pathways are modulated differently as evidenced by the heatmap. Such transcriptome profiling with a larger cohort can stratify patients into intrinsic subtypes and provide insight into the molecular mechanisms of podocyte injury.

Rice genes specifically expressed in a rice mutant gained resistance to rice blast.(oral)

  • C. U. Han;Lee, C. H.;K. S. Jang;Park, Y. H.;H. K. Lim;Kim, J.C.;Park, G. J.;J.S. Cha;Park, J. E.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.66.2-66
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    • 2003
  • A gain-of-function mutant, SHM-11 obtained through gamma-ray mutagenesis, is resistant to rice blast caused by Magnaporthe grisea while wild type Sanghaehyanghyella is highly susceptible to the same disease. The resistance in the mutant was not race-specific when we tested with four races (KJ-201, KI-1113a, KI-313, KI-409) of M. grisea. To identify genes involved disease resistance in the gain-of-function mutant, genes specifically expressed in the mutant were selected by suppression subtractive hybridization using cDNAS of blast-inoculated mutant and wild type as a tester and a driver, respectively, Random 200 clones from the subtracted library were selected and analyzed by DNA sequencing. The sequenced genes represented three major groups related with disease resistance; genes encoding PR proteins, genes probably for phytoalexin biosynthesis, and genes involved in disease resistance signal transduction. A gene encoding a putative receptor-like protein kinase was identified as highly expressed only in the gain-of-function mutant after blast infection. The role of the putative receptor-like protein kinase gene during blast resistance will be further studied.

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Antimicrobial Resistance Patterns and Resistance genes assay of Shigella sonnei Isolated in Korea for Five Years (최근 5년 동안 국내에서 분리된 Shigella sonnei의 항균제 내성 유형과 내성유전자형 분석)

  • Huh, Wan;Lee, Sang-Jo;Kwon, Gi-Seok;Jang, Jong-Ok;Lee, Jung-Bok
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.31-39
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    • 2007
  • This study has been carried out for investigating the relatedness of representative 135 Shigella sonnei strains isolated from 2000 to 2004 by using biotyping and antimicrobial resistance. All strains showed typical biochemical characterisics of Shigella strain. Among 135 strains,79 (58.5%) strains were biotype "g",54 (40.0%) strains were biotype "a" and 2 (1.5%) strains were biotype "e". The results of susceptibility test against 16 antimicrobial agents were like this. Most of strains were susceptible to AN, CIP, C and GM. 129 (95.6%) strains were resistant to SXT, 126 (93.3%) strains were resistant to TE and 122 (90.4%) strains were resistant to SM. One hundred thirty two (97.8%) strains were resistant to more than two antimicrobial agents. R28 type (antimicrobial resistance patterns 28: resistant to AM, SAM, TE, TIC, SXT, K, SM and AmC) were 42 strains (31.1%). The other strains were showed 33 kinds of R patterns. The results of $bla_{TEM}$, sulII, tetA and strA gene detection were coincided with phenotype of antimicrobial resistance by disk diffusion method. But some strains which had sulII and strA genes were not showed the resistance against SXT and SM.