• Animal Systematics, Evolution and Diversity
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• v.26 no.3
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• pp.183-186
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• 2010

To identify Korean red-backed voles (Myodes regulus) from Korea by mitochondrial DNA (mtDNA) sequencing, we obtained mtDNA control region sequences of 17 red-backed voles from Korea and northeast China, and these sequences were compared with the corresponding haplotypes of Myodes obtained from GenBank. We identified five red-backed voles from Mt. Changbai and Harbin as M. rufocanus and another three redbacked voles from Harbin as M. rutilus, respectively. Moreover, nine red-backed voles from Korea, showing the average nucleotide distance of 0.66% among nine haplotypes, were different from other species of Myodes, and the average distance between nine haplotypes of red-backed voles from Korea and seven haplotypes of M. rufocanus was 6.41%, whereas the average distance between nine haplotypes of red-backed voles from Korea and five haplotypes of M. rutilus was 14.8%. We identified the red-backed voles from Korea as M. regulus, and found that M. regulus is distinct in its mtDNA control region sequences as well, although we propose further analyses with additional specimens from East Asia using nuclear and mtDNA markers to confirm the distinctness of M. regulus.

• Genomics & Informatics
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• v.16 no.4
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• pp.23.1-23.5
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• 2018

Virus taxonomy was initially determined by clinical experiments based on phenotype. However, with the development of sequence analysis methods, genotype-based classification was also applied. With the development of genome sequence analysis technology, there is an increasing demand for virus taxonomy to be extended from in vivo and in vitro to in silico. In this study, we verified the consistency of the current International Committee on Taxonomy of Viruses taxonomy using an in silico approach, aiming to identify the specific sequence for each virus. We applied this approach to norovirus in Caliciviridae, which causes 90% of gastroenteritis cases worldwide. First, based on the dogma "protein structure determines its function," we hypothesized that the specific sequence can be identified by the specific structure. Firstly, we extracted the coding region (CDS). Secondly, the CDS protein sequences of each genus were annotated by the conserved domain database (CDD) search. Finally, the conserved domains of each genus in Caliciviridae are classified by RPS-BLAST with CDD. The analysis result is that Caliciviridae has sequences including RNA helicase in common. In case of Norovirus, Calicivirus coat protein C terminal and viral polyprotein N-terminal appears as a specific domain in Caliciviridae. It does not include in the other genera in Caliciviridae. If this method is utilized to detect specific conserved domains, it can be used as classification keywords based on protein functional structure. After determining the specific protein domains, the specific protein domain sequences would be converted to gene sequences. This sequences would be re-used one of viral bio-marks.

• Mycobiology
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• v.30 no.1
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• pp.1-4
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• 2002

Genetic relatedness of medically important Exophiala species such as E. dermatitidis, E. mansonii, and three E. jeanselmei varieties: jeanselmei, lecanii-corni, and heteromorpha was examined using PCR-RFLP(restriction fragment length polymorphism) of ribosomal DNA, M-13, $(GTG)_5$ and nucleotide sequences of ribosomal ITS(internal transcribed space) II regions. Three E. jeanselmei varieties showing distinct band patterns for each DNA markers as well as different nucleotide sequences of ribosomal ITS II regions could be considered as a separate species. E. dermatitidis and E. mansonii demonstrated the identical band patterns of RFLP of ribosomal DNA, M-13, and $(GTG)_5$ markers. However, nucleotides sequences of ribosomal ITS II region were different between these two species.

• Journal of Microbiology and Biotechnology
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• v.4 no.4
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• pp.274-277
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• 1994

We subcloned two chicken H3 histone genes and transfected them into Rat 3 cell line. One contains 300 bp 5' to its cap site and the other contains 130 bp 5' to its cap site when cloned into plasm ids. Both of them showed 5' phase specific expression of their mRNA about 8 fold higher (during 5' phase) than during Gl phase. This means that only 130 bp 5' to its cap site was enough to confer cell cycle regulated expression of the latter gene. The DNA sequences of their 5' flanking region did not reveal any particular homologies or subtype-specific sequences. The DNA sequence data also showed that even though the protein coding regions of the histone genes have been conserved exceptionally well throughout evolution, their 5' untranslated regions have not been conserved as well.

• Journal of Plant Biotechnology
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• v.40 no.1
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• pp.27-36
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• 2013

We isolated and functionally characterized a Dendrobium Actin1 (DmACT1) promoter that drives strong gene expression in the orchid genus Dendrobium. A genomic fragment containing the region 3227 bp upstream of the coding region of DmACT1 was obtained by inverse PCR. Detailed comparison of the full-length cDNA and genomic sequences revealed that DmACT1 has a 1374 bp first intron in the 5' UTR. However, the 5' flanking sequences upstream of the coding region showed no obvious sequence similarities compared to those of known promoters, including plant actin promoters. Serial deletion constructs of the 5' flanking region from the translation initiation codon were fused to the coding sequence of a GUS/luciferase fusion reporter to identify the regulatory elements necessary for promoter activity. Transient assays in the flowers of Dendrobium revealed that the 5' UTR-intron greatly enhanced promoter activity. Moreover, the DmACT1 promoter with its 5' UTR-intron yielded approximately 10-fold higher reporter activity than the rice Act1 promoter-intron. Our data suggest that the DmACT1 promoter with its 5' UTR-intron is a useful tool for strong expression of transgenes in Dendrobium orchids.

• Parasites, Hosts and Diseases
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• v.55 no.3
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• pp.279-285
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• 2017

The present study was performed to analyze molecularly the phylogenetic positions of human-infecting Trichostrongylus species in Mazandaran Province, Iran, which is an endemic area for trichostrongyliasis. DNA from 7 Trichostrongylus infected stool samples were extracted by using in-house (IH) method. PCR amplification of ITS2-rDNA region was performed, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 5.0 software. Six out of 7 isolates had high similarity with Trichostrongylus colubriformis, while the other one showed high homology with Trichostrongylus axei registered in GenBank reference sequences. Intra-specific variations within isolates of T. colubriformis and T. axei amounted to 0-1.8% and 0-0.6%, respectively. Trichostrongylus species obtained in the present study were in a cluster with the relevant reference sequences from previous studies. BLAST analysis indicated that there was 100% homology among all 6 ITS2 sequences of T. colubriformis in the present study and most previously registered sequences of T. colubriformis from human, sheep, and goat isolates from Iran and also human isolates from Laos, Thailand, and France. The ITS2 sequence of T. axei exhibited 99.4% homology with the human isolate of T. axei from Thailand, sheep isolates from New Zealand and Iran, and cattle isolate from USA.

• Mycobiology
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• v.38 no.1
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• pp.17-25
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• 2010

The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

• Journal of Mushroom
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• v.12 no.4
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• pp.251-257
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• 2014

The aim of this study was to analyze the genetic diversity of Hericium species based on their rDNA ITS sequences. Hericium species were collected from various regions and the size of the ITS rRNA gene regions from different Hericium species varied from 450 to 500 bp. A phylogenetic trees based on the ITS region revealed that Hericium species could be classified into 4 different groups, H. erinaceus, H. coralloide, H. alpestre, H. americanum. Among them, ASI 48015 and ASI 48016 was identified as Sprassis and Lentinula genus, respectively, based on blast searches using their rDNA ITS sequences.

• Korean Journal of Medicinal Crop Science
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• v.17 no.3
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• pp.165-172
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• 2009

The bramble cultivated widely in South Korea, which is known as Bokbunja-ddal-gi, is regarded as having originated from Korean native Rubus coreanus. This study was carried out to obtain basic phylogenetic information on Korean cultivated bramble (KCB) by comparing the internal transcribed spacer (ITS) regions with those of R. coreanus, blackberry (R. lanciniatus), black (R. occidentalis) and red (R. idaeus) raspberry. Sequences of the ITS 1 suggest that some KCB accessions share a significant similarity with both R. occidentalis and R. coreanus in the ITS 1 region. The ITS 2 sequences of the three KCB accessions clustered more closely to those of two R. occidentalis accessions than to those of R. coreanus. These results suggest that there exist variations in the sequences of ITS among KCB accessions and KCB accessions are more closely related to black raspberry than R. coreanus in the ITS regions.

• Korean Journal of Plant Taxonomy
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• v.36 no.4
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• pp.263-277
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• 2006

Anemone pendulisepala, recently described as a new species, is distributed in sympatry with A. reflexa, A. amurensis, and A. raddeana at Mt. taebeark and Mt. Baekdu. Anemone pendulisepala was previously proposed to be a hybrid species between A. reflexa and A. amurensis becaue it displavs overlapping features with them in involucre shape, petiole length, sepal apex and xylem shape, To verify the taxonomic status and to examine the hybridity of A. pendulisepala, sequences of ITS region of nuclear ribosomal DNA and the psba-trnH, rps16 and trnLF region of cpDNA from 36 accessions of 5 taxa including outgroup were analyzed. In maximum parsimony tree based on ITS sequences, A. pendulisepala had the same sequences of A. reflexa and was clustered with monophyletic A. amurensis, and then A. raddeana. Anemone pendulisepala was distinguished from the other taxa by having four base insertion in rps16 region, two species-specific bases and insertion in trnLF region. In the phylogenetic trees of combined cpDNA, A. pendulisepala showed monophyly with the bootstrap 100%. Anemone pendulisepala exhibited no polymorphism and shared no sequences with putative parental or related taxa examined in this study. Molecular data suggest that A. pendulisepala should be a distinct species, and no evidence of the hybridization was detectcd.