• Title/Summary/Keyword: gene diversity

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Phylogenetic Study of Genus Haliotis in Korea by Cytochrome c Oxidase Subunit 1 and RAPD Analysis (Cytochrome c oxidase subunit 1과 RAPD 분석에 의한 한국 전복속의 계통 연구)

  • Seo, Yong Bae;Kang, Sung Chul;Choi, Seong Seok;Lee, Jong Kyu;Jeong, Tae Hyug;Lim, Han Kyu;Kim, Gun-Do
    • Journal of Life Science
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    • v.26 no.4
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    • pp.406-413
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    • 2016
  • Abalones are gastropod mollusks belonging to the genus Haliotis. Pacific abalones are regarded as a very important marine gastropod mollusk in Korea, Japan, China, and also in food industries around the world. In Korea, 6 species of abalone have been reported to occur along the coasts: Haliotis discus hannai, Haliotis discus discus, Haliotis madaka, Haliotis gigantea, Haliotis diversicolor supertexta, and Haliotis diversicolor diversicolor. This study was performed to discriminate the genetic variances by the partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) genes and random amplified polymorphic DNA (RAPD) analysis against four species of Pacific abalone (H. discus hannai, H. discus, H. madaka, H. gigantea). COI gene is reasonably well conserved and has been sequenced in various invertebrate taxa. The RAPD analysis technique is a relatively simple and low cost method that allows differentiation of taxa without the need to know their genomes. In this study, we investigated the genetic diversity, phylogenetic relationships within each species. The COI and RAPD analysis were able to distinguish between H. gigantea and the other three species. However, these analysis methods were inadequate to distinguish between H. discus and H. madaka. These results are believed to be able to provide a basis data for future hybrid breeding research by defining the genetically closely related four species of abalone, which is to develop new hybrid abalone for export using hybrid breeding.

Analysis of the Diversity of Methicillin Resistant Staphylococcus aureus (MRSA) Strains Isolated from Diarrhea Patients in Busan Area (부산지역 설사환자에서 분리한 MRSA 균주의 다양성 분석)

  • Choi, Seong-Hwa;Park, Eun-Hee;Park, Yon-Koung;Kim, Joung-A;Kim, Nam-Ho;Lee, Young-Suk;Bin, Jae-Hun;Park, Ho-Kuk
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1083-1089
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    • 2008
  • We investigated the molecular epidemiological characteristic of methicillin resistant Staphylococcus aureus (MRSA) strains isolated from stool samples in Busan from 2004 to 2006. Among 142 isolates of S. aureus, 49 isolates (34.5%) were confirmed as MRSA. With the antimicrobial susceptibility tests, 37 isolates (75.5%) showed multiple resistance to more than 10 antibiotics, but all isolates were sensitive to vancomycin. All of MRSA had enterotoxin A in 30.6%, B 4.1%, C 8.3%, D, C/G, A 2.0% and None 51%. PFGE of SmaI-digested chromosomal DNA was performed on 49 sporadic MRSA isolates. Restriction fragment patterns consisted of 8 to 14 fragments ranged in size from 48.5 to 630.5 kbp. We could divided the isolates into 7 groups ($I{\sim}VII$) by analyzing PFGE patterns. Group I subdivided as 2 subgroups and 17 (34.7%) strains belong to the group I. Dendrogram of PFGE patterns showed that MRSA strains in Busan were heterogeneous but we could find out minor homogeneity in hospital.

Cloning and Expression of Partial Japanese Flounder (Paralichthys olivaceus) IgD

  • Choi, Dae-Han;Jang, Han-Na;Ha, Dae-Mang;Kim, Jae-Wha;Oh, Chan-Ho;Choi, Sang-Hoon
    • BMB Reports
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    • v.40 no.4
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    • pp.459-466
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    • 2007
  • The cDNA sequence of the Japanese flounder (Paralychthys olivaceus) IgD has been previously reported (GenBank accession no. AB052658) and this was followed by the detection of IgD mRNA expression in some flounder organ tissues. However, it has not been determined whether the flounder IgD gene is virtually expressed into IgD protein. To characterize the flounder immunoglobulins utilized in elucidating the mechanism, evolution and diversity of the flounder immune system, antibodies specific to IgD and IgM were necessary. In the present study, partial flounder recombinant IgD (rIgD), IgM (rIgM) and the conserved regions of IgD and IgM (rCIg) were produced by cloning the cDNA sequence using isotype specific primers which were designed to produce unique fragments of IgD and IgM specific amino acid sequences. The production of recombinant Igs was ascertained by SDS-gel electrophoresis and immunoblot analysis using anti-T7$\cdot}$Taq antibody. The produced recombinant Igs were purified using affinity columns, and used as immunogens. Antibodies specific to the isotype of flounder Igs were generated by immunizing rabbits with rfIgs and the antibodies produced were identified by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Specificities of the generated antibodies were evaluated by testing cross-reactivity between recombinant IgM and IgD. By ELISA, rabbit antibodies against the rfIgD fragment (anti-rfIgD) failed to recognize any kind of flounder serum Igs, whereas respective antibodies against rfCIg (anti-rfCIg) and rfIgM fragments (anti-rfIgM) reacted with serum Igs. Likewise, in immunoblot assays, though anti-rfIgD did not, both anti-rfCIg and anti-rfIgM bound with the ~85 kd flounder IgM heavy chain. By flow cytometry analysis, anti-rfCIg, anti-rfIgD and anti-rfIgM reacted with 6%, 3% and 6.5% of cells, respectively, suggesting that flounder IgD is not secreted in serum but expressed on flounder B-like cell surfaces as in mammals. Antibodies produced against recombinant flounder Igs could be used to develop sandwich assay systems for detecting flounder Igs and for further investigating the flounder immune system.

The Population Genetic Structure of the Japanese Anchovy (Engraulis japonicus Temminck & Schlegel) in the West, South and East Seas of Korea Based on Microsatellite DNA Analysis (Microsatellite을 이용한 서해, 남해 및 동해 멸치 계군 분석)

  • Oh, Taeg-Yun;Kim, Joo-Il;Seo, Young-Il;Cho, Eun-Seob
    • Journal of Life Science
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    • v.19 no.2
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    • pp.174-178
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    • 2009
  • The characteristics of the population genetic structure of the Japanese anchovy (Engraulis japonicus Temminck & Schlegel) were collected from the West, South and East Seas of Korea in August, 2006 and were compared using six microsatellite DNA loci. In the West Sea population, the range of allele number against 72 individuals was from 19 to 41, the average allele number was 28.5. In EJ9, the allele number had the highest value of 41, this was 1.4 times higher than the average number of allele. The average allele number of the South Sea population was 24.5 that was less than that of West Sea population. In EJ2, EJ9 and EJ27.1 loci, the allele number was higher than average allele number in the South Sea population. In the East Sea population, the average allele number was estimated at 25.0 that most of loci except for EJ35 were higher than average allele number. Allele frequency in the West, South and East Sea populations was below 0.24. The value of observed heterozosity for six loci was approximately 0.5 higher than that of expected heterozosity (p>0.05), but three populations similar values to these heterozosity. Although the genetic diversity was higher value of above 0.9, three populations had a similar value. Genetic differentiation and distance combined estimate of the six loci were 0.258 and 0.019 (p>0.05), respectively, but showed no significant distance between three populations. These results suggested that it is responsible for no differentiated gene pool between three populations.

Assessment of Genetic Diversity and Relationships Between Korean Cattle and Other Cattle Breeds by Microsatellite loci (Microsatellite loci 분석에 의한 한우와 타 품종간의 유전적 유연관계)

  • Yoon, D.H.;Park, E.W.;Lee, S.H.;Lee, H.K.;Oh, S.J.;Cheong, I.C.;Hong, K.C.
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.341-354
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    • 2005
  • For the genetic assessment of the cattle breeds including Hanwoo, eleven microsatellite markers on ten bovine autosomes were genetically characterized for 618 individuals of nineteen cattle breeds; North Eastern Asian breeds (Korean cattle, Korean Black cattle, Japanese Black cattle, Japanese Brown cattle, Yanbian cattle), Chinese yellow cattle (Luxi cattle, Nanyang cattle), European Bas taurus (Angus, Hereford, Charolais, Holstein, Limousin), African Bas taurus (N'Dama, Baoule), African Bas indicus (Kavirondo Zebu, White Fulani), Asian Bas indicus (Sahiwal, Nelore) and one Bali cattle, Bas banteng as an outbreed-reference population. Allele frequencies derived from the genotyping data were used in estimating heterozygosities, gene diversities and genetic distances. The microsatellite loci were highly polymorphic, with a total of 162 different alleles observed across all loci. Variability in allele numbers and frequencies was observed among the breeds. The average expected heterozygosity of North Eastern Asian breeds was higher than those of European and African taurines, but lower than those of Asian and African indicines. Genetic distances were estimated using Nei's DA genetic distance and the resultant DA matrix was used in the construction of the phylogenetic trees. The genetic distances between North Eastern Asian cattle breeds and Bas indicus were similar with those between European Bas taurus and Bas indicus, and African Bas taurus and Bas indicus, respectively. The clusters were clearly classified into North Eastern Asian, European and African taurines groups as well as different cluster with Chinese mainland breeds, firstly out-grouping with Bas indicus. These results suggest that Korean cattle, Hanwoo, had not been originated from a crossbred between Bas primigenius in Europe and Bas indicus in India and North Eastern Asian Bas taurus may be have separate domestication from European and African Bas taurus.

Bacterial Community Profiling during the Manufacturing Process of Traditional Soybean Paste by Pyrosequencing Method (Pyrosequencing을 이용한 전통된장 제조과정 중 세균군집구조의 분석)

  • Kim, Yong-Sang;Jeong, Do-Yeon;Hwang, Young-Tae;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.275-280
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    • 2011
  • In order to evaluate the diversity and change of bacterial population during the manufacturing process of traditional soybean paste (doenjang), bacterial communities were analyzed using 16S rRNA gene-based pyrosequencing. In rice straw, the most important inoculum source for fermentation, the bacterial sequences with a relative abundance greater than 1% were assigned to four phyla, Proteobacteria (71%), Actinobacteria (20.6%), Bacteroidetes (4.2%), and Firmicutes (1.3%). Unlike bacterial community composition of rice straw, a different pattern of bacterial population in meju was observed with predominantly high abundance (99.1%) of Firmicutes. Phylum composition in young doenjang was almost same as that of meju. Major genera in young doenjang were Bacillus (81.3%), Clostridium (6.9%) and Enterococcus (6.3%) and the predominant species among bacterial population was B. amyloliquefaciens (63.6%). Abundance of the phylum Firmicutes in mature doenjang was 99.98%, which was even higher value than those in meju and young doenjang. Predominant species in mature doenjang were B. amyloliquefaciens (67.3%), B. atrophaeus (12.7%), B. methylotrophicus (6.5%), B. mojavensis (3.2%), and B. subtilis. (2.5%), which were also identified as major species of the microbial flora in meju. These results suggested that rice straw was a primary source for supplement of Bacillus species in manufacturing the traditional doenjang and that some species of Bacillus strains were mainly involved in the fermentation process of traditional doenjang.

Screening and Isolation of a Gene Encoding 4-Hydroxyphenylpyruvate Dioxygenase from a Metagenomic Library of Soil DNA (토양의 DNA로부터 4-Hydroxyphenylpyruvate Dioxygenase 유전자 탐색 및 분리)

  • Yun, Sang-Soon;Lee, Jung-Han;Kim, Soo-Jin;Kim, Sam-Sun;Park, In-Cheol;Lee, Mi-Hye;Koo, Bon-Sung;Yoon, Sang-Hong;Yeo, Yun-Soo
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.345-351
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    • 2005
  • To access the natural products of uncultured microorganisms, we constructed and screened the metagenomic DNA libraries by using a cosmid vector and DNA inserts isolated directly from soil. Initial screening of the libraries in Escherichia coli resulted in the isolation of several clones that produce a dark brown color when grown in LB medium. One of the positive clones, designed pYS85C, was transposon mutagenized and the DNA surrounding the transposon insertions in cosmids that no longer conferred the production of brown pigment to E. coli was sequenced. Annotation of the pYS85C sequence obtained from the transposon mutagenesis experiment indicated a single 393 amino acid open reading frame (ORF) with a molecular mass of about 44.5 kDa, predicted to be a 4-hydroxyphenylpyruvate dioxygenases (HPPDs), was responsible for the observed brown pigment. In a BLAST search against deposited sequence, the translated protein from this ORF showed moderate-level identity (>60%) to the other known HPPDs and was most conserved in the C-terminal region of the protein. These results show that genes involved in natural product synthesis can be cloned directly from soil DNA and expressed in a heterologous host, supporting the idea that this technology has the potential to provide novel natural products from the wealth of environmental microbial diversity and is a potentially important new tool for drug discovery.

A report on 57 unrecorded bacterial species in Korea in the classes Betaproteobacteria and Gammaproteobacteria

  • Kim, Hyun Sik;Cha, Chang-Jun;Cho, Jang-Cheon;Im, Wan-Taek;Jahng, Kwang Yeop;Jeon, Che Ok;Joh, Kiseong;Kim, Seung Bum;Seong, Chi Nam;Kim, Wonyong;Yi, Hana;Lee, Soon Dong;Yoon, Jung-Hoon;Bae, Jin-Woo
    • Journal of Species Research
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    • v.6 no.2
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    • pp.101-118
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    • 2017
  • In an investigation of indigenous prokaryotic species in Korea, a total of 57 bacterial strains assigned to the classes Betaproteobacteria and Gammaproteobacteria were isolated from diverse environments. Samples were collected from fresh water, natural caves, soil, paddy fields, lakes, sea water, jeotgal (fermented seafood), salt flats, soil from abandoned mines, plant roots, digestive organs of both Japanese crested ibis (Nipponia nippon) and Burmese python (Python molurus bivittatus) and tidal flats. From the high 16S rRNA gene sequence similarity (>98.7%) and formation of robust phylogenetic clades with closely related species, it was determined that each strain belonged to an independent and predefined bacterial species within either the Betaproteobacteria or Gammaproteobacteria. There is no official report or publication that describes these 57 proteobacterial species in Korea. Overall, in the class Betaproteobacteria there were 16 species in 12 genera of 4 families in the order Burkholderiales and two species in two genera of one family in the order Neisseriales. Within the class Gammaproteobacteia, there were five species in four genera of four families in the order Alteromonadales, 12 species in 11 genera of one family in the order Enterobacteriales, four species in four genera of three families in the order Oceanospirillales, 11 species in four genera of two families in the order Pseudomonadales, two species in the order Vibrionales and five species in five genera of one family in the order Xanthomonadales. Gram reaction, colony and cell morphology, basic biochemical characteristics, isolation source and strain IDs are described in the species description section.

Conservative Genes of Less Orthologous Prokaryotes (Orthologs 수가 적은 원핵생물들의 보존적 유전자)

  • Lee, Dong-Geun
    • Journal of Life Science
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    • v.27 no.6
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    • pp.694-701
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    • 2017
  • Mycoplasma genitalium represents the smallest genome among mono-cultivable prokaryotes. To discover and compare the orthologs (conservative genes) among M. genitalium and 14 prokaryotes that are uncultivable and have less orthologs than M. genitalium, COG (clusters of orthologous groups of protein) analyses were applied. The analyzed prokaryotes were M. genitalium, one hyperthermophilic exosymbiotic archaeon Nanoarchaeum equitans, four intracellular plant pathogenic eubacteria of Candidatus Phytoplasma genus, and nine endosymbiotic eubacteria of phloem- and xylem-feeding insects. Among 367 orthologs of M. genitalium, 284 orthologs were conservative between M. genitalium and at least one other prokaryote. All 15 prokaryotes commonly have 29 orthologs, representing the significance of proteins in life. They belong to 25 translation-related, including 22 ribosomal proteins, 3 subunits of RNA polymerase, and 1 protein-folding-related. Among the 15 prokaryotes, 40 orthologs were only found in all four Candidatus Phytoplasma. The other nine Candidatus, all endosymbionts with insects, showed only a single common COG0539 (ribosomal protein S1), representing the diversity of orthologs among them. These results might provide clues to understand conservative genes in uncultivable prokaryotes, and may be helpful in industrial areas, such as handling prokaryotes producing amino acids and antibiotics, and as precursors of organic synthesis.

Correlation between Karyotype and Phenotype in Turner Syndrome (터너증후군의 핵형과 표현형간의 연관성)

  • Shim, Ye-Jee;Hwang, Young-Ju;Lee, Kun-Soo
    • Journal of Genetic Medicine
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    • v.6 no.1
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    • pp.67-73
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    • 2009
  • Purpose : In spite of the karyotype and phenotype diversity in Turner syndrome patients, there are few reports about such differences in Korea. We reviewed the data of chromosome abnormalities, clinical manifestations, and comorbidities of Turner syndrome patients in Kyungpook National University Hospital to compare them to the recent hypotheses about sex chromosome gene loci related to Turner symptoms. Materials and Methods : We identified the cytologic findings of 92 patients with Turner syndrome and the clinical findings of 62 patients among them. Results : 54.3 percent of patients had 45,X while 45.7 percent showed other karyotype combinations (45,X/46,XX, 45,X/46,XX/47,XXX, 46,X,del(Xp), 46,X,del(Xq), 45,X/46,X,del(Xq), 46,X,i(Xq), 45,X/46,X,i (Xq)). The Turner symptoms found included short neck, high arched palate, broad chest, Madelung deformity, short metacarpals, scoliosis, cubitus valgus, low hair line, webbed neck, edematous extremities, pigmented nevus, and sexual infantilism. The specific diseases associated Turner syndrome included renal abnormalities, congenital heart disease, hearing defects, diabetes mellitus, hyperlipidemia, and decreased bone density. The phenotype of the mosaicism group was milder than that of the monosomy group. In the case of 46,X,del(Xp) and 45,X/46,X,del(Xq) groups, all had skeletal abnormalities, but the 46,X,del(Xq) group had none. In the case of 46,X,del(Xp) group, all showed short statures and skeletal abnormalities, but no sexual infantilism was observed. In the case of 46,X,i(Xq) and 45,X/46,X,i(Xq) groups, they all showed delayed puberty and had primary amenorrhea. Conclusion : It is important to study karyotype-phenotype correlations in patients with Turner syndrome to obtain interesting information about the genotype-phenotype correlations related to the X chromosome.

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