• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.737-744
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• 2004

A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Vairimorpha spp. and Nosema spp. and identification of Vairimorpha necatrix from Lepidoptera insects. Three sets of primers were selected from different genomic sequences to specifically amplify an 831 bp amplicon within the SSU rRNA gene, specific for both Vairimorpha spp. and Nosema spp. (MSSR primer); a 542 bp amplicon within the SSU rRNA gene, specific for Vairimorpha spp. (VSSU primer); and a 476 bp amplicon within the actin gene, specific for Vairimorpha necatrix (VNAG primer). Using the primers in conjunction with multiplex PCR, it was possible to detect Vairimorpha spp. and Nosema spp. and to differentiate between them. The sensitivity of this PCR assay was approximately 10 spores per milliliter. It is proposed that the multiplex PCR is a sensitive, specific, and rapid tool that can serve as a useful differential diagnostic tool for detecting Vairimorpha spp. and Nosema spp. in Lepidoptera insect.

• Journal of the korean society of oceanography
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• 제39권3호
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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.

• Mycobiology
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• 제30권1호
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• pp.13-17
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• 2002

The genomic DNAs were extracted from roots of Glycine max and Sorghum bicolor, and compared with those from spores of two arbuscular mycorrhizal(AM) fungi, Glomus mosseae and Scutellospora heterogama. The partial small subunit(SSU) of ribosomal RNA genes were synthesized and amplified by polymerase chain reaction with the fungal specific primers, AM1 and NS31. By the recent molecular techniques, the presence of another AM fungal DNA were confirmed in the roots of two plants, and three sequences of rDNA fragments amplified were identified to be close to those of G. caledonium, G. fasiculatum and G. proliferum. The two AM fungi, both, were found to colonize at the cortical layers of plant roots collected in the fields, together.

• 생명과학회지
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• 제12권4호
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• pp.427-432
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• 2002

잇바디돌김(Porptyra dentata)을 대상으로 핵의 18S ribo-somal RNA를 지령하는 유전자 즉 18S rDNA 유전자를 증폭하고, 염기서열분석을 수행하였다. 전체 18S rDNA의 exon 영역 크기는 1822 bp, intron 영역의 크기는 512 bp였다. 이들 exon과 intron 영역의 G+C함량은 각각 49%와 55%씩 나타내었다. 일본산 잇바디돌김(CenBank accession number: AB013183)의 exon 영역과의 비교에서 상동성이 97.1%에 도달하였다. 568번과 569번 염기사이의 upstream에 위치하는 intron 영 역에서는 AB013183과 52.1%의 상동성을 보였다.

• Mycobiology
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• 제48권6호
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• pp.464-475
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• 2020

Acervus (Pyronemataceae, Pezizales) is a saprobic genus in Pezizomycetes, characterized by colored apothecia, subcylindrical to cylindrical asci and guttulate ascospores. We collected four Acervus samples from China and Thailand. Descriptions and illustrations are introduced for all fresh samples. One new record of A. globulosus from Thailand, one new species, A. rufus, two known species, A. epispartius and A. stipitatus from China are reported. Phylogenetic analysis based on five genes, the large subunit rRNA (LSU), the translation elongation factor-1 alpha (tef1-α), the second largest subunit of RNA polymerase II (rpb2), the largest subunit of RNA polymerase II (rpb1), and the small subunit rRNA (SSU), revealed the distinct position of the new species. The new species is set apart by its red apothecia. A key to Acervus species is also given.

• 생명과학회지
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• 제22권1호
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• pp.62-73
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• 2012

본 연구는 팽이버섯(Flammulina velutipes)의 국내 균주와 외래 균주간의 유전적 유연관계를 조사하기 위하여 수행되었으며, 계통수 측정 결과 3개 그룹으로 나눠짐을 확인하였다. 20개 F. velutipes 균주들의 ITS region 염기 서열을 확보하였으며, 이들 서열을 바탕으로 multiple alignment를 보았으며, Neighbor-joining method를 이용하여 계통수를 작성하였으며, 2개 그룹으로 나눠짐을 확인하였다. 또한 F. velutipes 4154 균주의 rDNA-cluster를 최초로 분석한 결과, 총 10,974 bp임을 밝혔다. SSU는 1,806 bp, ITS region은 553 bp의 염기배열이 결정되었다. ITS 1 부분은 245 bp이고 ITS 2는 308 bp였으며, LSU에 해당되는 염기서열은 3,402 bp, IGS 1은 1,400 bp, 5S는 83 bp, IGS 2는 3,571 bp임을 확인하였다.

• Parasites, Hosts and Diseases
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• 제54권4호
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• pp.423-429
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• 2016

Cryptosporidium is an important pathogen causing gastrointestinal disease in snakes and is distributed worldwide. The main objectives of this study were to detect and identify Cryptosporidium species in captive snakes from exotic pet shops and snake farms in Thailand. In total, 165 fecal samples were examined from 8 snake species, boa constrictor (Boa constrictor constrictor), corn snake (Elaphe guttata), ball python (Python regius), milk snake (Lampropeltis triangulum), king snake (Lampropeltis getula), rock python (Python sebae), rainbow boa (Epicrates cenchria), and carpet python (Morelia spilota). Cryptosporidium oocysts were examined using the dimethyl sulfoxide (DMSO)-modified acid-fast staining and a molecular method based on nested-PCR, PCR-RFLP analysis, and sequencing amplification of the SSU rRNA gene. DMSO-modified acid-fast staining revealed the presence of Cryptosporidium oocysts in 12 out of 165 (7.3%) samples, whereas PCR produced positive results in 40 (24.2%) samples. Molecular characterization indicated the presence of Cryptosporidium parvum (mouse genotype) as the most common species in 24 samples (60%) from 5 species of snake followed by Cryptosporidium serpentis in 9 samples (22.5%) from 2 species of snake and Cryptosporidium muris in 3 samples (7.5%) from P. regius.

• Parasites, Hosts and Diseases
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• 제60권1호
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• pp.45-49
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• 2022

The Japanese amberjack Seriolae quinqueradiata is one of the most consumed fish species among the Koreans. However, information regarding parasitic infection in Japanese amberjack is scarce. This study described the morphological and molecular characteristics of a species of philometrid nematode, Philometroides seriolae, which was recovered from Japanese amberjack. This fish was caught in the sea of Goseong-gun, Gangwon-do, Republic of Korea (Korea). Six P. seriolae (Nematoda: Philometridae) were recovered from 2 Japanese amberjacks. These parasites were subgravid female which were 325-420 mm long and 2.95-3.27 mm wide. Furthermore, they had typical papillae distributed on their body surface with 14 papillae at the apical view. Sequence analysis of the small subunits of ribosomal RNA (SSU rRNA) showed high sequence identity (99.8%, 1,607/1,611-bp) with that of P. seriolae (GenBank accession no. FJ155811). This nematode species has been newly added to the Korean nematode fauna.

• Asian-Australasian Journal of Animal Sciences
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• 제14권6호
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• pp.880-884
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• 2001

Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.

• Animal Systematics, Evolution and Diversity
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• 제29권2호
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• pp.144-151
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• 2013

Two marine hypotrichous ciliates, Anteholosticha petzi and Ponturostyla enigmatica, were collected from the Yellow Sea and the Korea Strait, respectively, and described using live observation and protargol-impregnated specimens. Furthermore, the nuclear small subunit ribosomal RNA gene of each was sequenced and compared to previously annotated sequences retrieved from the GenBank. Anteholosticha petzi is characterized by 3 frontal cirri (FC), 2 frontoterminal cirri (FTC), 8-12 transverse cirri (TC), 1 buccal cirrus (BC), 9-12 midventral pairs (MP), 3 bipolar dorsal kineties (DK), and 3 types of colorless cortical granules. Ponturostyla enigmatica is characterized by 8 FC, 5 ventral cirri (VC), 5-7 TC, 6-7 marginal rows (MR) on each side, 4 complete and 2-3 partial DK, and greenish cortical granules. This is the first identification and description of these 2 species, A. petzi and P. enigmatica, in South Korea.