• Title/Summary/Keyword: SSU1 gene

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Identical small subunit ribosomal RNA gene nucleotide sequence of bovine Theileria isolates (Korea and Japan) and Theileria buffeli (Marula, Kenya) (한국파 일본의 소에서 분리한 Theileria 분리주와 Theiferia buffeli (Marula, Kenya)의 small subunit ribosomal RNA 유전자 염기서열의 일치)

  • 채준석;권오덕
    • Parasites, Hosts and Diseases
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    • v.36 no.1
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    • pp.47-54
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    • 1998
  • Small subunit ribosomal RNA (SSU rRNA) gene nucleotide sequences of bovine ReiLerin isolates from Korea (KLS and KCB) and japan (JHS) were determined. The genes from each isolate were amplified by the polymerase chain reaction and the approxi- mately 1.8 kb product cloned and sequenced by a modified dideoxynucleotide method. Overlapping gene segments produced with a series of primers were sequenced, resoRting in a complete DNA sequence for both forward and reverse strands of the SSU rRNA genes of each isolate. SSU rRNA gene sequences (termed Type A) were identical among the bovine ReiLeri,n isolates from Korea and the isolate from Japan. A GenBank data library homolo- gy search showed the sequence to be the same as that listed as leiLeyia buKeLi isolated from cattle in Marula, Kenya.

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Three ORF-Containing Group I Introns in Chloroplast SSU of Caulerpa sertularioides (Ulvophyceae) and Their Evolutionary Implications

  • Lee, Jung-Ho;Manhart, James R.
    • ALGAE
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    • v.18 no.3
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    • pp.183-190
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    • 2003
  • Except for a group I intron in trnL-uaa occuring in eubacteria and plastids, group I introns are rarely documented in plastid genomes. Here, we report that a green alga, Caulerpa sertularioides, contains three group IA3 introns in the 16S gene (cpSSU), CS-cpSSU.i1, CS-cpSSU.i2 and CS-cpSSU.i3. Each intron has an open reading frame with LAGLIDADG motifs. CS-cpSSU.i1orf and CS-cpSSU.i3orf occur at Loop 6 in the intron secondary structure and CScpSSU. i2orf at Loop 8. CS-cpSSU.i1orf and CS-cpSSU.i2orf contain both LAGLI-DADG motifs but CS-cpSSU.i3orf has only one. CS-cpSSU.i1 and CS-cpSSU.i2 share the insetion sites and the ORFs at Loop 6 and 8 with CpSSU·1 and CpSSU·2 introns of Chlamydomonas pallidostigmatica (Chlorophyceae). In contrast, CS-cpSSU.i3, containing 28 copies of GAAATAT at Loop 6, is a novel intron found only in Caulerpa sertularioides. Possible scenarios of the evolution of the three introns and their possible use in systematic research are discussed.

PCR Diagnosis of Entamoeba histolytica Cysts in Stool Samples

  • Moon, Joung-Ho;Cho, Shin-Hyeong;Yu, Jae-Ran;Lee, Won-Ja;Cheun, Hyeng-Il
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.281-284
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    • 2011
  • Amebiasis is a protozoan disease caused by Entamoeba histolytica and a potential health threat in areas where sanitation and hygiene are inappropriate. Highly sensitive PCR methods for detection of E. histolytica in clinical and environmental samples are extremely useful to control amebiasis and to promote public health. The present study compared several primer sets for small subunit (SSU) rDNA and histone genes of E. histolytica cysts. A 246 bp of the SSU rDNA gene of pure cysts contained in phosphate-buffered saline (PBS) and in stool samples was successfully amplified by nested PCR, using the 1,147-246 bp primer set, of the primary PCR products which were pre-amplified using the 1,147 bp primer as the template. The detection limit of the nested PCR using the 1,147-246 primer set was 10 cysts in both groups (PBS and stool samples). The PCR to detect histone gene showed negative results. We propose that the nested PCR technique to detect SSU rDNA can be used as a highly sensitive genetic method to detect E. histolytica cysts in stool samples.

Molecular Phylogenetic Relationships Within the Genus Alexandrium(Dinophyceae) Based on the Nuclear-Encoded SSU and LSU rDNA D1-D2 Sequences

  • Kim, Choong-Jae;Sako Yoshihiko;Uchida Aritsune;Kim, Chang-Hoon
    • Journal of the korean society of oceanography
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    • v.39 no.3
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    • pp.172-185
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    • 2004
  • LSU rDNA D1-D2 and SSU rDNA genes of 23 strains in seven Alexandrium (Halim) species, A. tamarense (Lebour) Balech, A. catenella (Whedon et Kofoid), A. fraterculus (Balech) Balech, A. affine (Inoue et Fukuyo) Balech, A. insuetum Balech, A. pseudogonyaulax (Biecheler) Horiguchi ex Yuki et Fukuyo and A. tamiyavanichii Balech, were sequenced and the data were used for molecular phylogenetic analysis. The sequence data revealed 11 and 7 ribotypes in the LSU rDNA D1-D2 region and 4 and 17 ribotypes in the SSU rDNA region of A. catenella and A. tamarense, respectively. Other Alexandrium species had also 1 to 5 ribotypes in the two regions. With the exception of CMC2 and CMC3 of A. catenella, all A. tamarense and A. catenella strains had a common ribotype, a functionally expressed rRNA gene (here termed type A), in both gene regions. In addition to the functionally expressed gene, several pseudogenes were obtained that were found to be good tools to analyze the population designation of regional isolates by grouping them according to shared ribotypes. From the phylogenetic analysis of the sequence data determined in this study and retrieved from GenBank, the genus Alexandrium was divided into 14 groups: 1) A. tamarense, 2) A. excavatum, 3) A. catenella, 4) Tasmanian A. tamarense, 5) A. affine (and/or A. concavum), 6) Thai A. tamarense, 7) A. tamiyavanichii, 8) A. fraterculus, 9) A. margalefii, 10) A. andersonii, 11) A. ostenfeldii, 12) A. minutum (or A. lusitanicum), 13) A. insuetum, and 14) A. pseudogonyaulax. The SSU rDNA gene sequence of A. fundyense was so similar to those of A. tamarense used in this study that the two species were difficult to discriminate each other. A. tamiyavanichii was closest to the A. tamarense strain isolated in Thailand and close to the long chain-forming species of A. affine and A. fraterculus. The phylogenetic tree showed that A. margalefii, A. andersonii, A. ostenfeldii, A. minutum and A. insuetum constituted the basal relative complex, and that A. pseudogonyaulax is an ancestral taxon in the genus Alexandrium.

Cryptosporidium suis Infection in Post-Weaned and Adult Pigs in Shaanxi Province, Northwestern China

  • Lin, Qing;Wang, Xing-Ye;Chen, Jian-Wen;Ding, Ling;Zhao, Guang-Hui
    • Parasites, Hosts and Diseases
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    • v.53 no.1
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    • pp.113-117
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    • 2015
  • Cryptosporidium spp., ubiquitous enteric parasitic protozoa of vertebrates, recently emerged as an important cause of economic loss and zoonosis. The present study aimed to determine the distribution and species of Cryptosporidium in post-weaned and adult pigs in Shaanxi province, northwestern China. A total of 1,337 fresh fecal samples of post-weaned and adult pigs were collected by sterile disposable gloves from 8 areas of Shaanxi province. The samples were examined by Sheather's sugar flotation technique and microscopy at${\times}400$ magnification for Cryptosporidium infection, and the species in positive samples was further identified by PCR amplification of the small subunit (SSU) rRNA gene. A total of 44 fecal samples were successfully amplified by the nested PCR of the partial SSU rRNA, with overall prevalence of 3.3%. The average prevalence of Cryptosporidium infection in each pig farms ranged from 0 to 14.4%. Species identification by sequencing of SSU rRNA gene revealed that 42 (3.1%) samples were Cryptosporidium suis and 2 (0.15%) were Cryptosporidium scrofarum. C. suis had the highest prevalence (7.5%) in growers and the lowest in breeding pigs (0.97%). C. suis was the predominant species in pre-weaned and adult pigs, while C. scrofarum infected pigs older than 3 months only. A season-related difference of C. suis was observed in this study, with the highest prevalence in autumn (5.5%) and the lowest (1.7%) in winter. The present study provided basic information for control of Cryptosporidium infection in pigs and assessment of zoonotic transmission of pigs in Shaanxi province, China.

Cloning and Organization of the Ribosomal RNA Genes of the Mushroom Trichloma matsutake

  • Hwang, Seon-Kap;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.194-199
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    • 1995
  • A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).

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Phylogenetic relationships among Acanthamoeba spp. based on PCR-RFLP analyses of mitochondrial small subunit rRNA gene

  • Yu, Hak-Sun;Hwang, Mee-Yul;Kim, Tae-Olk;Yun, Ho-Cheol;Kim, Tae-Ho;Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.37 no.3
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    • pp.181-188
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    • 1999
  • We investigated the value of mitochondrial small subunit rRNA gene (mt SSU rDNA) PCR-RFLP as a taxonomic tool for Acanthamoeba isolates with close inter-relationships. Twenty-five isolates representing 20 species were included in the analysis. As in nuclear 18s rDNA analysis, two type strains (A. astronyxis and A. tubiashi) of morphological group 1 diverged earliest from the other strains, but the divergence between them was less than in 18s riboprinting. Acanthamoeba griffini of morhological group 2 branched between pathogenic (A. culbertsoni A-1 and A. healyi OC-3A) and nonpathogenic (A.palestinensis Reich, A. pustulosa GE-3a, A. royreba Oak Ridge, and A lenticulata PD2S) strains of morphological group 3. Among the remaining isolates of morphological group 2, the Chang strain had the identical mitochondrial riboprints as the type strain of A. hatchetti. AA2 and AA1, the type strains of A. divionensis and A. paradivionensis, respectively, had the identical riboprints as A. quina Vil3 and A. castellanii Ma. Although the branching orders of A. castellanii Neff, A. polyphaga P23, A. triangularis SH621, and A. lugdunensis L3a were different from those in 18S riboprinting analysis, the results obtained from this study generally coincided well with those from 18S riboprinting. Mitochondrial riboprinting may have an advantage over nuclear 18S rDNA riboprinting beacuse the mt SSU rDNAs do not seem to have introns that are found in the 18S genes of Acanthamoeba and that distort phylogenetic analyses.

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Novel Discovery of Two Heterotrichid Ciliates, Climacostomum virens and Fabrea salina (Ciliophora: Heterotrichea: Heterotrichida) in Korea

  • Kim, Ji Hye;Shin, Mann Kyoon
    • Animal Systematics, Evolution and Diversity
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    • v.31 no.3
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    • pp.182-190
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    • 2015
  • Two heterotrichid ciliates, Climacostomum virens (Ehrenberg, 1838) Stein, 1859 from brackish water and freshwater, and Fabrea salina Henneguy, 1890 from a solar saltern, were collected in Korea. They are novelly investigated in Korea by means of live observation, protargol staining and nuclear small subunit (SSU) rRNA gene sequencing. Climacostomum virens is characterized by pouch-like body shape, body length of $200-370{\mu}m$ in vivo, conspicuous cytopharyngeal tube, macronuclei ribbon-like shape, and one to four in number, with or without symbiont algae in cytoplasm, 34-66 somatic kineties, 67-113 adoral zone of membranelles, 8-42 peristomial kineties, 24-37 apical membranelles. SSU rDNA sequence size is 1,591 bp and GC contents 48.52%. Fabrea salina is also characterized by scoop-like body shape with proboscis, body length of $190-240{\mu}m$ in vivo, one to two rod-shaped macronuclei, oval micronuclei, grayish green cortical granules, 104-186 somatic kineties, 4-8 preoral kineties, 7-19 peristomial kineties and fragmented paroral membrane. SSU rDNA sequence size is 1,598 bp and GC contents 47.50%.

The Wine Yeast Strain-Dependent Expression of Genes Implicated in Sulfide Production in Response to Nitrogen Availability

  • Mendes-Ferreira, A.;Barbosa, C.;Jimenez-Marti, E.;Del Olmo, M.;Mendes-Faia, A.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.9
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    • pp.1314-1321
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    • 2010
  • Sulfur metabolism in S. cerevisiae is well established, but the mechanisms underlying the formation of sulfide remain obscure. Here, we investigated by real-time RT-PCR the dependence of expression levels of MET3, MET5/ECM17, MET10, MET16, and MET17 along with SSU1 on nitrogen availability in two wine yeast strains that produce divergent sulfide profiles. MET3 was the most highly expressed of the genes studied in strain PYCC4072, and SSU1 in strain UCD522. The strains behaved differently according to the sampling times, with UCD522 and PYCC4072 showing the highest expression levels at 120 h and 72 h, respectively. In the presence of 267 mg assimilable N/l, the genes were more highly expressed in strain UCD522 than in PYCC4072. MET5/ECM17 and MET17 were only weakly expressed in both strains under any condition tested. MET10 and SSU1 in both strains, but MET16 only in PYCC4072, were consistently upregulated when sulfide production was inhibited. This study illustrates that strain genotype could be important in determining enzyme activities and therefore the rate of sulfide liberation. This linkage, for some yeast strains, of sulfide production to expression levels of genes associated with sulfate assimilation and sulfur amino acid biosynthesis could be relevant for defining new strategies for the genetic improvement of wine yeasts.

Redescription of Two Marine Ciliates (Ciliophora: Urostylida: Pseudokeronopsidae), Pseudokeronopsis carnea and Uroleptopsis citrina, from Korea

  • Baek, Ye-Seul;Jung, Jae-Ho;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.27 no.3
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    • pp.220-227
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    • 2011
  • The morphology of the two marine urostyloid ciliates, Pseudokeronopsis carnea (Cohn, 1866) and Uroleptopsis citrina Kahl, 1932, in the family Pseudokeronopsidae, collected from the Yellow Sea, and the East Sea, Korea, respectively, were studied using live observation and protargol impregnation. Additionally, the small subunit ribosomal RNA (SSU rRNA) gene was sequenced. These two species are firstly recorded in Korea. The main diagnostic key is as follows. Pseudokeronopsis carnea: body outline elongate-elliptical, brown-reddish or orange-red in colour in vivo; bicorona of 16-24 frontal cirri; one buccal and two frontoterminal cirri; 7-10 transverse cirri; 5-7 dorsal kineties; two types of cortical granules (one orange-red pigment, mainly grouped around cirri and dorsal bristles, arranged in typical rubra-pattern; the other, colourless and blood-cell-shaped, and densely distributed); contractile vacuole in the posterior half of the cell on the left side, usually in posterior 1/3-2/5. Uroleptopsis citrina: body outline elongate-elliptical, lemon-yellow in colour in vivo; two types of cortical granules (one yellow pigment; the other, blood-cell-shaped, densely distributed); bicorona of 12-18 frontal cirri; 2-3 frontoterminal cirri; two midventral rows comprising 26-35 cirri (consisting of anterior paired cirri, non-paired single cirri, and posterior paired cirri); three dorsal kineties. In addition, the SSU rRNA sequences of the two species were compared with public database of these species and consequently, showed high similarity.