• Title/Summary/Keyword: intergenic region

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Rapid Detection of the pathogenic agent of Bacterial white enteritis of Larval and Juvenile Stages in Olive flounder (Paralichthys olivaceus) (넙치(Paralichthys olivaceus)자치어 장관백탁증(Bacterial white enteritis) 원인균의 신속 검출)

  • Mun, Yeong-Geon;Park, Geun-Tae;Son, Hong-Ju;Lee, Sang-Hyeon;Lee, Jeong-Min;Heo, Mun-Su
    • Journal of fish pathology
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    • v.17 no.3
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    • pp.159-169
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    • 2004
  • Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

Vibrio 4종의 16s-23s ribosomal DNA Intergenic Spacer region의 유전학적 분석

  • 장인권;이실한;이승환;최용석;임현정;송재희
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.163-164
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    • 2000
  • Vibrio는 새우양식장 및 종묘배양장의 사육수에서 가장 qlsqjsgkrp 출현하며 양식새우의 질병과 직간접적으로 관련을 갖고 있는 주요 병원성 세균류로서 이들 종의 신속한 동정은 질병의 조기진단 및 대책을 위하여 매우 중요하기 때문에 이에 대한 연구가 많이 이루어져 왔으나 일부 Vibrio 종들간의 생리, 화학적 특성이 너무도 유사하여 동정에 많은 어려움을 겪고 있다. (중략)

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16S/23S Intergenic Spacer Region as a Genetic Marker for Thiobacillus thiooxidans and T.ferrooxidans

  • Lee, Hye-Won;Choi, Won-Young;Cho, Kyung-Suk;Choi, Won-Ja
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1046-1054
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    • 2001
  • Bioleaching is the process in which insoluble metal sulfide is oxidized by specialized iron- and/or sulfur-oxidizing lithotrophic bacteria in acidic, metal-rich environments. Most of these processes are carried out by the genus Thiobacillus. Three novel Thiobacillus strains (Thiobacillus thiooxidans AZ11, Thiobacillus thiooxidans MET, and thiobacillus thiooxidans TAS) associated with bioleaching have been isolated from soil and sludge (Korean patent No. 1999-0073060 for T. thiooxidans AZ11, Korean patent No. 1999-0005798 for T. thiooxidans MET, and Korean patent No. 1999-0073059 for T. thiooxidans TAS). A partial sequence of 16S ribosomal RNA gene (16S rDNA) and the entire sequence of 16S/23S intergenic spacer region (ISR) were determined in the three above novel strains and in Thiobacillus ferrooxidans ATCC19859 as a reference strain. When phylogenetic analysis was performed based on G+C contents and sequence alignments, T. ferroxidans ATCC19859 was found to be closely related to previously registered T. ferrooxidans strains in a monophyletic manner, while the three novel T. thiooxidans strains were classified in a paraphyletic manner. Close examination on the base composition of 16S/23S ISR revealed that the 5\` part (nucleotide residues 21-200) was specific for the genus Thiobacillus. On the other end, the 3\` part (nucleotide residues 201-520) showed specificity in T. ferrooxidans strains, but not in T. thiooxidans strains. These results suggest that the proximal and distal halves of 16S/23S could be used as a genetic marker for the identification of the genus Thiobacillus and the species T. ferrooxidans, respectively.

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Genome-Wide Association Study of Orthostatic Hypotension and Supine-Standing Blood Pressure Changes in Two Korean Populations

  • Hong, Kyung-Won;Kim, Sung Soo;Kim, Yeonjung
    • Genomics & Informatics
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    • v.11 no.3
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    • pp.129-134
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    • 2013
  • Orthostatic hypotension (OH) is defined by a 20-mm Hg difference of systolic blood pressure (dtSBP) and/or a 10-mm Hg difference of diastolic blood pressure (dtDBP) between supine and standing, and OH is associated with a failure of the cardiovascular reflex to maintain blood pressure on standing from a supine position. To understand the underlying genetic factors for OH traits (OH, dtSBP, and dtDBP), genome-wide association studies (GWASs) using 333,651 single nucleotide polymorphisms (SNPs) were conducted separately for two population-based cohorts, Ansung (n = 3,173) and Ansan (n = 3,255). We identified 8 SNPs (5 SNPs for dtSBP and 3 SNPs for dtDBP) that were repeatedly associated in both the Ansung and Ansan cohorts and had p-values of < $1{\times}10^{-5}$ in the meta-analysis. Unfortunately, the SNPs of the OH case control GWAS did not pass our p-value criteria. Four of 8 SNPs were located in the intergenic region of chromosome 2, and the nearest gene (CTNNA2) was located at 1 Mb of distance. CTNNA2 is a linker between cadherin adhesion receptors and the actin cytoskeleton and is essential for stabilizing dendritic spines in rodent hippocampal neurons. Although there is no report about the function in blood pressure regulation, hippocampal neurons interact primarily with the autonomic nervous system and might be related to OH. The remaining SNPs, rs7098785 of dtSBP trait and rs6892553, rs16887217, and rs4959677 of dtDBP trait were located in the PIK3AP1 intron, ACTBL2-3' flanking, STAR intron, and intergenic region, respectively, but there was no clear functional link to blood pressure regulation.

Cooperative transcriptional activation of ATP-binding cassette sterol transporters ABCG5 and ABCG8 genes by nuclear receptors including Liver-X-Receptor

  • Back, Su Sun;Kim, Jinsu;Choi, Daehyung;Lee, Eui Sup;Choi, Soo Young;Han, Kyuhyung
    • BMB Reports
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    • v.46 no.6
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    • pp.322-327
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    • 2013
  • The ATP-binding cassette transporters ABCG5 and ABCG8 form heterodimers that limit absorption of dietary sterols in the intestine and promote cholesterol elimination from the body through hepatobiliary secretion. To identify cis-regulatory elements of the two genes, we have cloned and analyzed twenty-three evolutionary conserved region (ECR) fragments using the CMV-luciferase reporter system in HepG2 cells. Two ECRs were found to be responsive to the Liver-X-Receptor (LXR). Through elaborate deletion studies, regions containing putative LXREs were identified and the binding of $LXR{\alpha}$ was demonstrated by EMSA and ChIP assay. When the LXREs were inserted upstream of the intergenic promoter, synergistic activation by $LXR{\alpha}/RXR{\alpha}$ in combination with GATA4, $HNF4{\alpha}$, and LRH-1, which had been shown to bind to the intergenic region, was observed. In conclusion, we have identified two LXREs in ABCG5/ABCG8 genes for the first time and propose that these LXREs, especially in the ECR20, play major roles in regulating these genes.

Determining Potential Link between Environmental and Clinical Isolates of Cryptococcus neoformans/Cryptococcus gattii Species Complexes Using Phenotypic and Genotypic Characterisation

  • Kenosi Kebabonye;Mosimanegape Jongman;Daniel Loeto;Sikhulile Moyo;Wonderful Choga;Ishmael Kasvosve
    • Mycobiology
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    • v.51 no.6
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    • pp.452-462
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    • 2023
  • Opportunistic infections due to Cryptococcus neoformans and C. gattii species complexes continue to rise unabated among HIV/AIDS patients, despite improved antifungal therapies. Here, we collected a total of 20 environmental and 25 presumptive clinical cryptococcal isolates from cerebrospinal fluid (CSF) samples of 175 patients enrolled in an ongoing clinical trial Ambition 1 Project (Botswana-Harvard Partnership). Identity confirmation of the isolates was done using MALDI-TOF MS and PCR. We describe the diversity of the isolates by PCR fingerprinting and sequencing (Oxford Nanopore Technology) of the intergenic spacer region. Mating types of the isolates were determined by amplification of the MAT locus. We report an unusual prevalence of 42.1% of C. neoformans × C. deneoformans hybrids Serotype AD (n = 16), followed by 39.5% of C. neoformans Serotype A (n = 15), 5.3% of C. deneoformans, Serotype D (n = 2), 7.9% of C. gattii (n = 3), and 5.3% of C. tetragattii (n = 2) in 38 representative isolates that have been characterized. Mating type-specific PCR performed on 38 representative environmental and clinical isolates revealed that 16 (42.1%) were MATa/MAT𝛼 hybrids, 17 (44.7%) were MAT𝛼, and five (13.2%) possessed MATa mating type. We used conventional and NGS platforms to demonstrate a potential link between environmental and clinical isolates and lay a foundation to further describe mating patterns/history in Botswana.

Comparison of scanning electron microscopic structures and nucleotide sequences variation of ITS1, 5.8S ribosomal RNA gene and ITS2 region in three Peruvian entomopathogenic fungal isolates (3종의 페루산 entomopathogenic fungi의 전자현미경적 구조와 ITS1, 5.8S ribosomal RNA gene, ITS2의 염기서열 다양성)

  • Han, Sang-Hoon;Nam, Sunghee;Lee, Heui-Sam;Yeo, Joo-Hong
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.137-141
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    • 2013
  • In this study, nucleotide sequence structures of intergenic transcribed spacer (ITS) 1, complete 5.8S ribosomal RNA gene and ITS 2 region were analyzed to identify three Peruvian entomopathogenic fungal isolates. The isolates had highly conserved sequence region in 5.8S rRNA gene and unique sequences in ITS 1 and 2 region among them. 5.8S rRNA gene regions were highly conserved and showed high homoloies among tested isolates. In contrast, ITS region showed species-specific sequence region, resulting in inter-genus differencies. Scanning electron microscopic images of these isolates supported the result of ITS-based identification. From these result, Peruvian entomopathogenic fungal isolate J270, J278, were identified as Beauveria bassiana and J271 was identified as Lecanicillium attenuatum.

Phylogenetic Relationships of Korean Viola (Violaceae) Based on matK and atpB-rbcL Sequence Data of Chloroplast DNA (엽록체 DNA의 matK와 aptB-rbcL 염기서열 분석에 의한 제비꽃속(Viola)의 계통유연관계)

  • Yoo, Ki-Oug;Jang, Su-Kil;Lee, Woo-Tchul
    • Korean Journal of Plant Taxonomy
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    • v.37 no.1
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    • pp.1-15
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    • 2007
  • Phylogenetic studies were conducted for 42 populations of Korean viola based on matK gene and atpB-rbcL intergenic spacer region of chloroplast DNA. In the matK tree, section Chamaemelanium and Dischidium were formed as a distinct group. Five subsections of section Nomimium were paraphyletic. In atpB-rbcL intergenic spacer region analysis, two species of sect. Chamaemelanium were monophyletic, and section Dischidium was placed sister to subsection patellares clade except for V. keiskei. Five subsections of section Nomimium were also paraphyletic as matK tree. the separate data analyses were incongruent in the relationships among 42 populations, especially for the position of section Dischidium and V. keiskei. The combined analyses of two chloroplast regions showed three major clades; section Chamaemelanium and Dischidium (x=6) formed a sister to subsections Hypocarpae and Trigonocarpae (x=10) clade; subsections Bilobatae and vaginatae (x=10 or 12) formed a clade with V. keiskei; and 19 populations of subsection patellares (x=12) except for V. keiskei were recognized as an independent clade within section Nomimium. Although combined data suggest three major clades of Korean viola, the origins of each clade from outgroup were discordance with previous ITS and trnL-F data.

Phylogenetic relationships of Coryloideae based on waxy and atpB-rbcL sequences (Waxy와 atpB-rbcL 염기서열 분석에 의한 Coryloideae의 계통 유연관계)

  • Yoo, Ki-Oug;Wen, Jun
    • Korean Journal of Plant Taxonomy
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    • v.38 no.4
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    • pp.371-388
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    • 2008
  • Phylogenetic studies were conducted for 35 populations of the subfamily Coryloideae (Betulaceae) based on waxy gene of nuclear DNA and atpB-rbcL intergenic spacer region of chloroplast DNA. Waxy data analysis suggest that Coryloideae is monophyletic; Corylus is monophyletic and basally branching within the subfamily Coryloideae; Ostryopsis is sister to the Carpinus and Ostrya clade, and the Ostrya is monophyletic (BS=86, PP=99). AtpB-rbcL intergenic spacer region analysis shows that Ostryopsis appeared as the most basal clade within the Coryloideae; Corylus is monophyletic(BS=98, PP=100) and placed between Carpinus-Ostrya and Ostryopsis clade; Carpinus and Ostrya formed a clade with a high support value(BS=100, PP=100). Carpinus sect. Carpinus is monophyletic, whereas sect. Distegocarpus is paraphyletic in the waxy tree. Corylus formed two subclades, but discordance at the infrageneric classification based on morphological characters. In the atpB-rbcL tree, Carpinus and Corylus taxa form a polytomy within the each clade. Results from the two data sets differ mainly in the relative position of Ostryopsis, the monophyly of Ostrya, and the relationships within the Carpinus-Ostrya clade. Further studies are needed for clarify the taxonomic position and the generic limitation.

Genetic Variation in Fusarium oxysporum f. sp. fagariae Populations Based RAPD and rDNA RFLP Analyses

  • Nagaraian, Gopal;Nam, Myeong-Hyeon;Song, Jeong-Young;Yoo, Sung-Joon;Kim, Hong-Gi
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.264-270
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    • 2004
  • Fusarium oxysporum f. sp. fragariae is a fungal pathogen causing strawberry wilt disease. The random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphisms (RFLPs) of intergenic spacer (IGS) region of rDNA were used to identify genetic variation among 22 F. oxysporum f. sp. fragariae isolates. All isolates could be distinguished from each other by RAPD analysis and RFLP of 2.6 kb amplified with primer CNS1 and CNL12 for IGS region of rDNA. Cluster analysis using UPGMA showed eight distinct clusters based on the banding patterns obtained from RAPD and rDNA RFLP. These results indicate that F. oxysporum f. sp. fragariae isolates are genetically distinct from each other, There was a high level genetic variation among F. oxysporum f. sp. fragariae.